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[Molecular characteristics of Japanese encephalitis virus carried by Culex tritaeniorhynchus in Dongchuan District, Kunming City, Yunnan Province]. [云南省昆明市东川区三带喙库蚊携带日本脑炎病毒的分子特征]。
Q3 Medicine Pub Date : 2024-07-18 DOI: 10.16250/j.32.1374.2023237
Y Gu, Y He, Y Chen, Z Yang, N Li, S Lü, Y Zhu, F Ruan, J Wang, J Wang
<p><strong>Objective: </strong>To isolate the Japanese encephalitis virus carried by <i>Culex tritaeniorhynchus</i> in Dongchuan District of Yunnan Province and analyze its molecular characteristics, so as to provide insights into the prevention and control of Japanese encephalitis in Yunnan Province.</p><p><strong>Methods: </strong>Mosquito specimens were collected using mosquito-trapping lamps from pig farms in Batang Village and Xiaoxin Village, Dongchuan District, Kunming City, Yunnan Province in July 2016, and the mosquito species was identified according to the mosquito morphology. Then, 60 to 100 mosquitoes of each species served as a group and were ground. Baby hamster kidney-21 (BHK-21) cells and <i>Aedes albopictus</i> clone C6/36 cells were used for virus isolation, and positive isolates were identified using flavivirus primers. The positive isolates were amplified using reverse transcription polymerase chain reaction (RT-PCR) assay with 15 pairs of specific primers covering the full length of the genotype I Japanese encephalitis virus, and DNA sequence assembly was performed using the software SeqMan in the DNASTAR package. The obtained sequences were aligned with the complete sequences of 38 Japanese encephalitis virus downloaded from the GenBank with the software MegAlign, and the nucleotide and amino acid homology analyses of the obtained sequences were performed. The difference in amino acid sites was analyzed with the software GeneDoc, and phylogenetic trees were created based on the sequences of the coding region and E protein of the isolated Japanese encephalitis virus with the software Mega X. In addition, the secondary and tertiary structures of the E protein of the Japanese encephalitis virus were predicted using the online tool SOPMA and the software Swiss-Model.</p><p><strong>Results: </strong>A total of 5 820 mosquitoes were collected and 3 843 <i>Cx. tritaeniorhynchus</i> (66.03%) were identified according to the mosquito morphology. A positive virus isolate, termed YNDC55-33, was isolated from <i>Cx. tritaeniorhynchoides</i> following batches of virus isolation from mosquito specimens, and cytopathic effect was observed following inoculation into BHK-21 and C6/36 cells. The YNDC55-33 virus isolate was successfully amplified with the flavivirus primes, and a long sequence containing 300 nucleotides was obtained. Following sequence alignment using the BLAST tool, the sequence of the YNDC55-33 virus isolate had high homology with that of the genotype I Japanese encephalitis virus. A long sequence with 10 845 nucleotides in length, which encoded 3 432 amino acids, was obtained by splicing the full sequence of the YNDC55-33 virus isolate. Phylogenetic analysis based on the whole-genome sequence and <i>E</i> gene sequence of the YNDC55-33 virus isolate showed that the new YNDC55-33 virus isolate was most closely related to the genotype I Guizhou isolate (GenBank accession number: HM366552), with nucleotide homology of 98.5% a
目的分离云南省东川区三带喙库蚊携带的日本脑炎病毒,分析其分子特征,为云南省日本脑炎的防控提供参考:2016年7月在云南省昆明市东川区八塘村和小新村的养猪场用诱蚊灯采集蚊子标本,根据蚊子形态鉴定蚊子种类。然后,以每种蚊子 60 至 100 只为一组,进行研磨。用小仓鼠肾-21(BHK-21)细胞和白纹伊蚊克隆C6/36细胞进行病毒分离,并用黄病毒引物鉴定阳性分离株。用 15 对特异性引物(覆盖基因型 I 日本脑炎病毒的全长)进行反转录聚合酶链反应(RT-PCR),扩增阳性分离物,并用 DNASTAR 软件包中的 SeqMan 软件进行 DNA 序列组装。用 MegAlign 软件将获得的序列与从 GenBank 下载的 38 个日本脑炎病毒的完整序列进行比对,并对获得的序列进行核苷酸和氨基酸同源性分析。此外,还利用在线工具 SOPMA 和软件 Swiss-Model 预测了日本脑炎病毒 E 蛋白的二级和三级结构:结果:共收集了 5 820 只蚊子,根据蚊子的形态鉴定出 3 843 只三线蓟马(66.03%)。在从蚊子标本中分批分离病毒后,从三带喙蚊中分离出了一个阳性病毒分离物,称为 YNDC55-33,接种到 BHK-21 和 C6/36 细胞后观察到细胞病理效应。YNDC55-33 病毒分离物经黄病毒原核扩增成功,获得了包含 300 个核苷酸的长序列。使用 BLAST 工具进行序列比对后,YNDC55-33 病毒分离物的序列与基因型 I 日本脑炎病毒的序列具有高度同源性。通过拼接 YNDC55-33 病毒分离株的全序列,得到了一个长度为 10 845 个核苷酸的长序列,该序列编码 3 432 个氨基酸。基于 YNDC55-33 病毒分离株全基因组序列和 E 基因序列的系统进化分析表明,新的 YNDC55-33 病毒分离株与基因 I 型贵州分离株(GenBank 编号:HM366552)的亲缘关系最密切,核苷酸同源性为 98.5%,氨基酸同源性为 99.4%,YNDC55-33病毒分离株与其他基因型I日本脑炎病毒分离株的核苷酸同源性为97.96%±0.33%,氨基酸同源性为99.35%±0.08%,与其他基因型日本脑炎病毒的核苷酸同源性<90%,氨基酸同源性<98%。YNDC55-33病毒分离株与候选减毒活疫苗SA14-14-2分离株在E基因上有16个氨基酸位点不同,在与神经毒性相关的8个关键氨基酸位点中有7个不同。据预测,YNDC55-33 病毒分离株 E 蛋白的二级和三级结构以随机线圈为特征:结论:从昆明市东川区三带喙蝮蛇中分离到了一种基因I型日本脑炎病毒。该病毒分离株与减毒活疫苗候选株SA14-14-2在抗原表位相关的关键氨基酸位点上无差异,病毒分离株的主要蛋白结构为随机线圈。这项研究为日本脑炎病毒在云南省的流行病学分布增添了新的数据,可为云南省日本脑炎的防控提供启示。
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引用次数: 0
[Surveillance of parasitic infections in market-sold aquatic products and knowledge and practice towards food-borne parasitic diseases among residents in Shanghai Municipality from 2020 to 2023]. [2020 - 2023年上海市市售水产品寄生虫感染监测及居民食源性寄生虫病防治知识与实践]。
Q3 Medicine Pub Date : 2024-07-08 DOI: 10.16250/j.32.1374.2024021
S Dai, Q Yu, Z Wang, Y Zhang, J Yang, Z Wei, X Ma, Q Zhu, J Chen, H Pan, H Wu
<p><strong>Objective: </strong>To investigate the prevalence of parasitic infections in market-sold aquatic products in Shanghai Municipality, and to understand the knowledge and practice towards food-borne parasitic diseases among residents, so as to provide insights into the surveillance and control of food-borne parasitic diseases.</p><p><strong>Methods: </strong>Freshwater products, seawater products and pickled products were randomly obtained from agricultural trade markets, supermarkets, retail stores and restaurants in Huangpu, Putuo, Minhang and Qingpu districts of Shanghai Municipality from 2020 to 2023. Parasite metacercariae and larvae were detected in these aquatic products using pressing method, digestion method and the dissection method, and the detection of parasitic infection was compared in different types of aquatic products. In addition, the knowledge and practice towards food-borne parasitic diseases were investigated among residents aged 10-80 years old who randomly selected from agricultural trade markets, supermarkets, community streets and other population concentration areas in these four districts by questionnaire in 2023, and the awareness of food-borne parasitic diseases knowledge and percentage of healthy behaviors formation were analyzed.</p><p><strong>Results: </strong>A total of 2 109 aquatic products sampled from Huangpu, Putuo, Minhang and Qingpu districts of Shanghai Municipality from 2020 to 2023 were detected, and there were 317 products detected with parasitic infections, with an overall detection rate of 15.03%. There were 8 products detected with parasitic infections in 1 221 freshwater products, with a detection rate of 0.66%, and <i>Clonorchis sinensis</i> was the predominant parasite, which was detected in <i>Pseudorasbora parva</i>, <i>Rhodeus amarus</i> and <i>Carassius auratus</i>. There were 82 products detected with parasitic infections in 501 seawater products, with a detection rate of 16.37%, and <i>Anisakis</i> was the predominant parasite, which was detected in <i>Trichiurus lepturus</i>, <i>Larimichthys polyactis</i>, <i>Pneumatophorus japonicus</i>, <i>Collchthys lucidus</i>, <i>Mugil cephalus</i>, <i>Larimichthys crocea</i>, <i>Scomberomorus niphonius</i>, <i>Stromateoides argenteus</i> and <i>Cololabis saira</i>. There were 227 products detected with parasitic infections in 387 pickled products, with a detection rate of 58.66%, and the prevalence rates of <i>Echinostoma metacercariae</i> were 76.27% (225/295) in <i>Bullacta exarata</i> and 11.11% (2/18) in crab pastes, respectively (<i>χ</i><sup>2</sup> = 159.511, <i>P</i> < 0.05). No <i>Paragonimus</i> infection was found in freshwater shrimps, crabs and pickled products, and no <i>Diphyllobothrium latum</i> larvae infection was detected in freshwater and seawater fish. Questionnaire surveys showed that the awareness of food-borne parasitic diseases knowledge was 79.00% (222/281), and the percentages of washing hands before and after meals,
目的:调查上海市市售水产品中寄生虫感染的流行情况,了解居民食源性寄生虫病的知识和做法,为食源性寄生虫病的监测和控制提供依据。方法:随机抽取2020 - 2023年上海市黄浦区、普陀区、闵行区和青浦区农贸市场、超市、零售商店和食肆的淡水产品、海水产品和腌制产品。采用挤压法、消化法和解剖法对水产品中寄生虫囊蚴和幼虫进行检测,并比较不同类型水产品中寄生虫感染的检测情况。并于2023年通过问卷调查的方式,随机抽取4区农贸市场、超市、社区街道等人口集中地区10-80岁居民对食源性寄生虫病的知识和行为进行调查,分析食源性寄生虫病知识知晓率和健康行为形成率。结果:2020 - 2023年,上海市黄埔、普陀、闵行、青浦区共检出2 109份水产品,检出寄生虫感染317份,总检出率为15.03%。在1 221份淡水产品中检出寄生虫感染8种,检出率为0.66%,以华支睾吸虫为优势寄生虫,分别在小伪螺旋体、大菱鲆和鲫鱼中检出。501种海水产品共检出寄生虫感染82种,检出率为16.37%,以异尖线虫为优势寄生虫,检出的主要品种有:细毛癣菌、多角鳞鱼、日本肺蝗、透明斑蝗、头毛癣菌、大角鳞鱼、尼松鳞鱼、银纹层蛾和沙蚤。387种腌渍产品中检出227种产品存在寄生虫感染,检出率为58.66%,白颡鱼和蟹膏中囊蚴检出率分别为76.27%(225/295)和11.11%(2/18),差异有统计学意义(χ2 = 159.511, P < 0.05)。淡水虾、蟹和腌制制品中未检出吸虫感染,淡水和海水鱼类中未检出叶状双芽绦虫幼虫感染。问卷调查显示,食源性寄生虫病知识知知率为79.00%(222/281),饭前饭后洗手、不直接饮用过滤水或自来水、不吃生的或半生的食物、愿意改变特殊饮食偏好或不良生活习惯以保持健康、愿意了解更多食源性寄生虫病知识的比例分别为99.64%(280/281)、72.24%(203/281)、56.23%(158/281)。黄浦区、普陀区、闵行区和青浦区分别占96.80%(272/281)和97.51%(274/281)。结论:上海市市售水产品存在食源性寄生虫污染。虽然居民了解食源性寄生虫病的知识,但需要对食源性寄生虫病进行可持续监测并改进健康教育,以尽量减少人类寄生虫感染的风险。
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引用次数: 0
[Advances in detection methods for fish-borne parasites in aquatic products]. [水产品中鱼类寄生虫检测方法的进展]。
Q3 Medicine Pub Date : 2024-07-04 DOI: 10.16250/j.32.1374.2024049
Y Zhang, Z Wang, J Chen, Q Yu

Consumption of raw and semi-raw aquatic products is strongly associated with the development of fish-borne parasitic diseases. Detection of fish-borne parasites in aquatic products is of great significance for the prevention and control of fish-borne parasitic diseases. This review describes the advances in the application of etiological, molecular biological and immunological techniques alone and in combinations for detection of fish-borne parasites, so as to provide insights into detection of fish-borne parasites.

食用生食和半生食水产品与鱼源性寄生虫病的发生密切相关。检测水产品中的鱼源性寄生虫对预防和控制鱼源性寄生虫病具有重要意义。本综述介绍了单独或联合应用病原学、分子生物学和免疫学技术检测鱼源性寄生虫的进展,从而为检测鱼源性寄生虫提供启示。
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引用次数: 0
[Epidemiological characteristics of visceral leishmaniasis in Gansu Province from 2017 to 2023]. [2017-2023年甘肃省内脏利什曼病流行病学特征]。
Q3 Medicine Pub Date : 2024-07-04 DOI: 10.16250/j.32.1374.2024018
F Li, D Yu, H Liang, C Yang, G Yang, J Yang

Objective: To analyze the epidemiological characteristics of visceral leishmaniasis in Gansu Province from 2017 to 2023, so as to provide insights into formulation of the visceral leishmaniasis control strategy in the province.

Methods: All epidemiological features of confirmed and clinically diagnosed cases of visceral leishmaniasis reported in Gansu Province from January 1, 2017 to December 31, 2023 were retrieved from the Chinese Disease Prevention and Control Information System, and the epidemiological characteristics of visceral leishmaniasis cases were analyzed descriptively.

Results: A total of 280 visceral leishmaniasis cases were reported in 39 counties (cities and districts) of Gansu Province from 2017 to 2023, including 258 local cases reported in 21 endemic counties (districts) and 22 imported cases reported in 18 non-endemic areas. Of the 280 cases, there were 262 cases with mountain type zoonotic visceral leishmaniasis (MT-ZVL), 12 cases with desert-type zoonotic visceral leishmaniasis (DT-ZVL), and 6 cases with unknown type. Re-emerging MT-ZVL occurred in Maiji District, Qinzhou District, Lixian County, Kangxian County, Zhenyuan County, Qin'an County and Yongjing County, and re-emerging DT-ZVL occurred in Dunhuang City, while emerging DT-ZVL occurred in Yumen City. The five counties (districts) reporting the highest number of visceral leishmaniasis cases included Wudu District, Wenxian County, Tanchang County, Zhouqu County and Diebu County, and a total of 220 cases were reported in these five counties, accounting for 78.57% of all visceral leishmaniasis cases in the province. Visceral leishmaniasis cases were reported each month throughout the year, with the peak in July. All reported visceral leishmaniasis cases had ages of 6 months to 81 years, including 50.71% (142/280) under 15 years of age, 49.29% (138/280) at ages of 15 years and older, and of all cases under 15 years of age, children at ages of 0 to 3 years were the most commonly affected (27.14%, 76/280). Among 280 visceral leishmaniasis cases, there were 173 males and 107 females with a male to female ratio of 1.62∶1, and farmer was the most common occupation (40.36%), followed by diaspora children (37.86%).

Conclusions: The prevalence of visceral leishmaniasis appeared an overall tendency towards a decline in Gansu Province from 2017 to 2023; however, there are still multiple challenges for visceral leishmaniasis control in the province. Reinforced dog monitoring and management, intensified human health education and improved capability building among professionals are recommended to manage the rebounding and spread of visceral leishmaniasis.

目的分析2017年至2023年甘肃省内脏利什曼病流行病学特征,为制定该省内脏利什曼病控制策略提供参考:从中国疾病预防控制信息系统中检索2017年1月1日至2023年12月31日甘肃省报告的所有内脏利什曼病确诊病例和临床诊断病例的流行病学特征,对内脏利什曼病病例的流行病学特征进行描述性分析:2017年至2023年甘肃省39个县(市、区)共报告内脏利什曼病病例280例,其中21个流行县(区)报告本地病例258例,18个非流行区报告输入病例22例。280例病例中,山地型人畜共患内脏利什曼病(MT-ZVL)262例,荒漠型人畜共患内脏利什曼病(DT-ZVL)12例,未知类型6例。麦积区、秦州区、礼县、康县、镇原县、秦安县和永靖县发生重发MT-ZVL,敦煌市发生重发DT-ZVL,玉门市发生新发DT-ZVL。报告内脏利什曼病病例数最多的5个县(区)包括武都区、文县、宕昌县、舟曲县和迭部县,这5个县共报告病例220例,占全省内脏利什曼病病例总数的78.57%。全年每月都有内脏利什曼病病例报告,7 月份是高峰期。所有报告的内脏利什曼病病例的年龄在 6 个月至 81 岁之间,其中 15 岁以下的占 50.71%(142/280),15 岁及以上的占 49.29%(138/280),而在所有 15 岁以下的病例中,0 至 3 岁的儿童最常见(27.14%,76/280)。在 280 例内脏利什曼病病例中,男性 173 例,女性 107 例,男女比例为 1.62∶1,农民是最常见的职业(40.36%),其次是散居儿童(37.86%):2017年至2023年,甘肃省内脏利什曼病流行率总体呈下降趋势,但该省内脏利什曼病防控仍面临多重挑战。建议加强犬只监测管理,强化人类健康教育,提高专业人员能力建设,以控制内脏利什曼病的反弹和蔓延。
{"title":"[Epidemiological characteristics of visceral leishmaniasis in Gansu Province from 2017 to 2023].","authors":"F Li, D Yu, H Liang, C Yang, G Yang, J Yang","doi":"10.16250/j.32.1374.2024018","DOIUrl":"https://doi.org/10.16250/j.32.1374.2024018","url":null,"abstract":"<p><strong>Objective: </strong>To analyze the epidemiological characteristics of visceral leishmaniasis in Gansu Province from 2017 to 2023, so as to provide insights into formulation of the visceral leishmaniasis control strategy in the province.</p><p><strong>Methods: </strong>All epidemiological features of confirmed and clinically diagnosed cases of visceral leishmaniasis reported in Gansu Province from January 1, 2017 to December 31, 2023 were retrieved from the Chinese Disease Prevention and Control Information System, and the epidemiological characteristics of visceral leishmaniasis cases were analyzed descriptively.</p><p><strong>Results: </strong>A total of 280 visceral leishmaniasis cases were reported in 39 counties (cities and districts) of Gansu Province from 2017 to 2023, including 258 local cases reported in 21 endemic counties (districts) and 22 imported cases reported in 18 non-endemic areas. Of the 280 cases, there were 262 cases with mountain type zoonotic visceral leishmaniasis (MT-ZVL), 12 cases with desert-type zoonotic visceral leishmaniasis (DT-ZVL), and 6 cases with unknown type. Re-emerging MT-ZVL occurred in Maiji District, Qinzhou District, Lixian County, Kangxian County, Zhenyuan County, Qin'an County and Yongjing County, and re-emerging DT-ZVL occurred in Dunhuang City, while emerging DT-ZVL occurred in Yumen City. The five counties (districts) reporting the highest number of visceral leishmaniasis cases included Wudu District, Wenxian County, Tanchang County, Zhouqu County and Diebu County, and a total of 220 cases were reported in these five counties, accounting for 78.57% of all visceral leishmaniasis cases in the province. Visceral leishmaniasis cases were reported each month throughout the year, with the peak in July. All reported visceral leishmaniasis cases had ages of 6 months to 81 years, including 50.71% (142/280) under 15 years of age, 49.29% (138/280) at ages of 15 years and older, and of all cases under 15 years of age, children at ages of 0 to 3 years were the most commonly affected (27.14%, 76/280). Among 280 visceral leishmaniasis cases, there were 173 males and 107 females with a male to female ratio of 1.62∶1, and farmer was the most common occupation (40.36%), followed by diaspora children (37.86%).</p><p><strong>Conclusions: </strong>The prevalence of visceral leishmaniasis appeared an overall tendency towards a decline in Gansu Province from 2017 to 2023; however, there are still multiple challenges for visceral leishmaniasis control in the province. Reinforced dog monitoring and management, intensified human health education and improved capability building among professionals are recommended to manage the rebounding and spread of visceral leishmaniasis.</p>","PeriodicalId":38874,"journal":{"name":"中国血吸虫病防治杂志","volume":"36 4","pages":"339-345"},"PeriodicalIF":0.0,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142355893","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Prevalence and genetic characteristics of Cryptosporidium infections among HIV-positive individuals in Jiangxi Province]. 江西省hiv阳性人群隐孢子虫感染流行及遗传特征分析
Q3 Medicine Pub Date : 2024-07-01 DOI: 10.16250/j.32.1374.2024071
Z Hu, L Lu, Y Yu, L Li, W Wang, G Fan, C Feng, Y Zheng, G Peng

Objective: To investigate the prevalence of Cryptosporidium infection and the distribution of parasite species and genotypes among HIV-positive individuals in Jiangxi Province.

Methods: HIV-positive individuals' sociodemographic and clinical data were collected from three AIDS designated hospitals in Jiangxi Province from January 2022 to March 2023. Subjects' stool samples were collected, and genomic DNA was extracted from stool samples. Nested PCR assay was performed based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium, and Cryptosporidium gp60 gene was amplified in stool samples positive for the SSU rRNA gene. The second-round PCR amplification product was checked with 1.5% agarose gel electrophoresis, and the products of suspected positive amplifications were sequenced, followed by sequence alignment. The phylogenetic tree was created using the Neighbor-Joining method with the software MEGA 11.0, to characterize the species, genotypes and sub-genotypes of Cryptosporidium.

Results: A total of 382 HIV-positive individuals were enrolled, with two cases identified with Cryptosporidium infection (0.52% prevalence), and both cases had no abdominal pain or diarrhea. Following sequencing and sequence alignment, the gene sequences of these two Cryptosporidium isolates shared 99.76% and 99.88% similarity with the gene sequence of C. meleagridis isolates. Phylogenetic analysis based on the Cryptosporidium SSU rRNA gene sequence identified the species of these two Cryptosporidium-positive stool samples as C. meleagridis. Following nested PCR amplification of the Cryptosporidium gp60 gene, sequencing and sequence alignment, the two C. meleagridis isolates were characterized as III eA17G2R1 and III bA25G1R1a sub-genotypes, and the sub-genotype III bA25G1R1a was firstly described in humans.

Conclusions: The prevalence of Cryptosporidium is low among HIV-positive individuals in Jiangxi Province. The likelihood of Cryptosporidium infection cannot be neglected among HIV-positive individuals without diarrhea.

目的:了解江西省hiv阳性人群隐孢子虫感染情况、寄生虫种类及基因型分布。方法:收集江西省3家艾滋病定点医院2022年1月至2023年3月hiv阳性个体的社会人口学和临床资料。收集受试者的粪便样本,并从粪便样本中提取基因组DNA。基于隐孢子虫小亚单位核糖体RNA (SSU rRNA)基因进行巢式PCR检测,在SSU rRNA基因阳性的粪便样本中扩增隐孢子虫gp60基因。第二轮PCR扩增产物经1.5%琼脂糖凝胶电泳检测,可疑阳性扩增产物测序,比对序列。采用MEGA 11.0软件,采用Neighbor-Joining法建立系统发育树,对隐孢子虫的种类、基因型和亚基因型进行表征。结果:共纳入382例hiv阳性个体,其中2例鉴定为隐孢子虫感染(患病率0.52%),均无腹痛和腹泻。经测序比对,两株隐孢子虫分离株的基因序列与C. meleagridis分离株的基因序列相似性分别为99.76%和99.88%。基于隐孢子虫SSU rRNA基因序列的系统发育分析,鉴定这两份隐孢子虫阳性粪便标本的种为隐孢子虫。对隐孢子虫gp60基因进行巢式PCR扩增、测序和序列比对,鉴定为III eA17G2R1和III bA25G1R1a亚基因型,其中III bA25G1R1a亚基因型首次在人类中发现。结论:江西省hiv阳性人群隐孢子虫感染率较低。在没有腹泻的hiv阳性个体中,隐孢子虫感染的可能性不容忽视。
{"title":"[Prevalence and genetic characteristics of <i>Cryptosporidium</i> infections among HIV-positive individuals in Jiangxi Province].","authors":"Z Hu, L Lu, Y Yu, L Li, W Wang, G Fan, C Feng, Y Zheng, G Peng","doi":"10.16250/j.32.1374.2024071","DOIUrl":"https://doi.org/10.16250/j.32.1374.2024071","url":null,"abstract":"<p><strong>Objective: </strong>To investigate the prevalence of <i>Cryptosporidium</i> infection and the distribution of parasite species and genotypes among HIV-positive individuals in Jiangxi Province.</p><p><strong>Methods: </strong>HIV-positive individuals' sociodemographic and clinical data were collected from three AIDS designated hospitals in Jiangxi Province from January 2022 to March 2023. Subjects' stool samples were collected, and genomic DNA was extracted from stool samples. Nested PCR assay was performed based on the small subunit ribosomal RNA (<i>SSU rRNA</i>) gene of <i>Cryptosporidium</i>, and <i>Cryptosporidium gp60</i> gene was amplified in stool samples positive for the <i>SSU rRNA</i> gene. The second-round PCR amplification product was checked with 1.5% agarose gel electrophoresis, and the products of suspected positive amplifications were sequenced, followed by sequence alignment. The phylogenetic tree was created using the Neighbor-Joining method with the software MEGA 11.0, to characterize the species, genotypes and sub-genotypes of <i>Cryptosporidium</i>.</p><p><strong>Results: </strong>A total of 382 HIV-positive individuals were enrolled, with two cases identified with <i>Cryptosporidium</i> infection (0.52% prevalence), and both cases had no abdominal pain or diarrhea. Following sequencing and sequence alignment, the gene sequences of these two <i>Cryptosporidium</i> isolates shared 99.76% and 99.88% similarity with the gene sequence of <i>C. meleagridis</i> isolates. Phylogenetic analysis based on the <i>Cryptosporidium SSU rRNA</i> gene sequence identified the species of these two <i>Cryptosporidium</i>-positive stool samples as <i>C. meleagridis</i>. Following nested PCR amplification of the <i>Cryptosporidium gp60</i> gene, sequencing and sequence alignment, the two <i>C. meleagridis</i> isolates were characterized as III eA17G2R1 and III bA25G1R1a sub-genotypes, and the sub-genotype III bA25G1R1a was firstly described in humans.</p><p><strong>Conclusions: </strong>The prevalence of <i>Cryptosporidium</i> is low among HIV-positive individuals in Jiangxi Province. The likelihood of <i>Cryptosporidium</i> infection cannot be neglected among HIV-positive individuals without diarrhea.</p>","PeriodicalId":38874,"journal":{"name":"中国血吸虫病防治杂志","volume":"36 6","pages":"637-642"},"PeriodicalIF":0.0,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143013401","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Development and preliminary application of a multiplex PCR assay for simultaneous detection of four intestinal parasites in goats]. [用于同时检测山羊体内四种肠道寄生虫的多重 PCR 检测方法的开发和初步应用]。
Q3 Medicine Pub Date : 2024-06-20 DOI: 10.16250/j.32.1374.2024046
Y Li, X Mu, H Xu, X Luo, R Yu, X Xu, L Yang, X Yu, Y Hong
<p><strong>Objective: </strong>To develop a multiplex PCR assay for simultaneous detection of four intestinal parasites, including <i>Giardia duodenalis</i>, <i>Cryptosporidium parvum</i>, <i>Enterocytozoon bieneusi</i> and <i>Moniezia</i>, and to preliminarily evaluate its detection efficiency.</p><p><strong>Methods: </strong>Four pairs of specific primers were designed based on the conserved sequences of the corresponding genes of <i>G. duodenalis</i> (GenBank accession number: XM_001710026.2), <i>C. parvum</i> (GenBank accession number: XM_626998.1), <i>E. bieneusi</i> (GenBank accession number: KJ719492.1) and <i>Moniezia</i> (GenBank accession number: OM296991.1) retrieved from the GenBank database, and a multiplex PCR assay for simultaneous detection of <i>G. duodenalis</i>, <i>C. parvum</i>, <i>E. bieneusi</i> and <i>Moniezia</i> was developed and optimized. A total of 116 fresh goat stool samples were collected from four goat farms in Zhanjiang City, Guangdong Province during the period from October to December 2022, including 96 samples used for evaluating the detection efficacy of the multiplex PCR assay, and 20 samples as baseline controls for sample testing. Genomic DNA extracted from 96 goat stool samples was tested using the single-target PCR assay and the developed multiplex PCR assay, and the sensitivity, specificity, positive predictive value, and negative predictive value of the multiplex PCR assay were evaluated for detection of <i>G. duodenalis</i>, <i>C. parvum</i>, <i>E. bieneusi</i> and <i>Moniezia</i> DNA in goat stool samples with the single-target PCR assay as the gold standard.</p><p><strong>Results: </strong>The multiplex PCR assay developed in this study allowed simultaneous amplification of specific gene fragments of <i>G. duodenalis</i>, <i>C. parvum</i>, <i>E. bieneusi</i> and <i>Moniezia</i>, with 1 400, 755, 314 bp and 585 bp in sizes, respectively, and the detection limit was 10<sup>2</sup> and higher copies of parasite DNA clones, while the multiplex PCR assay was negative for gene amplification of <i>Schistosoma japonicum</i>, <i>Fasciola hepatica</i>, <i>Echinococcus granulosus</i>, <i>Blastocystis hominis</i> and <i>Homalogaster paloniae</i>. Single-target PCR assay and the developed multiplex PCR assay were employed to test DNA samples extracted from 96 goat stool samples, and single-target PCR assay tested positive in 40 goat stool samples (41.67%), including 39 positive samples tested with the multiplex PCR assay, with a mean coincidence rate of 97.50% (39/40). The multiplex PCR assay tested positive for <i>G. duodenalis</i> DNA in 26 goat stool samples (27.10%), <i>C. parvum</i> DNA in 22 samples (22.90%), <i>E. bieneusi</i> DNA in 24 samples (25.00%), and <i>Moniezia</i> in 9 samples (9.40%), which was consistent with the detection using the single-target PCR assay. The sensitivity, negative predictive value, and positive predictive value of the multiplex PCR assay were 96.15%, 95.83%, 100.00% and 100.0
目的方法:根据十二指肠贾第虫(G. duodenalis)(GenBank登录号:XM_001710026.2)、副隐孢子虫(Cryptosporidium parvum)、生物肠虫(Enterocytozoon bieneusi)和莫尼茨虫(Moniezia)的相应基因的保守序列,设计了四对特异性引物:根据十二指肠孢子虫(GenBank登录号:XM_001710026.2)、副隐孢子虫(GenBank登录号:XM_626998.1)、肠孢子虫(GenBank登录号:KJ719492.1) 和莫尼茨绦虫(GenBank登录号:OM296991.1),并开发和优化了同时检测十二指肠球菌、副猪嗜血杆菌、E. bieneusi 和莫尼茨绦虫的多重 PCR 检测方法。在2022年10月至12月期间,从广东省湛江市的4个山羊养殖场共采集了116份新鲜山羊粪便样本,其中96份样本用于评估多重PCR检测方法的检测效果,20份样本作为基线对照进行样本检测。用单靶标PCR检测法和所开发的多重PCR检测法检测96份山羊粪便样本中提取的基因组DNA,并以单靶标PCR检测法为金标准,评估了多重PCR检测法检测山羊粪便样本中十二指肠球菌、副粘病毒、双球菌和莫尼茨绦虫DNA的灵敏度、特异性、阳性预测值和阴性预测值:结果:本研究开发的多重 PCR 检测法可同时扩增十二指肠球菌、副嗜血杆菌、嗜血杆菌和莫尼茨绦虫的特定基因片段。而多重 PCR 法对日本血吸虫、肝片吸虫、粒状棘球蚴、人型布氏囊虫和巴氏杆菌的基因扩增呈阴性。采用单目标 PCR 检测法和所开发的多重 PCR 检测法检测了从 96 份山羊粪便样本中提取的 DNA 样本,单目标 PCR 检测法检测了 40 份山羊粪便样本(41.67%)呈阳性,其中用多重 PCR 检测法检测了 39 份阳性样本,平均吻合率为 97.50%(39/40)。多重 PCR 检测法检测到 26 份山羊粪便样本(27.10%)中的十二指肠球虫 DNA 呈阳性,22 份样本(22.90%)中的副猪嗜血杆菌 DNA 呈阳性,24 份样本(25.00%)中的生物大肠杆菌 DNA 呈阳性,9 份样本(9.40%)中的莫尼茨虫 DNA 呈阳性,这与单靶标 PCR 检测法的检测结果一致。多重 PCR 检测法的灵敏度、阴性预测值和阳性预测值分别为 96.15%、95.83%、100.00% 和 100.00%、98.90%、98.92%、100.00% 和 100.00%、100.00%、100.00%、100.如果将单靶点 PCR 检测作为金标准,则山羊粪便样本中的 G. duodenalis、C. parvum、E. bieneusi 和 Moniezia DNA 的检测灵敏度分别为 98.90%、98.92%、100.00%、100.00% 和 100.00%:结论:已开发出一种高灵敏度和特异性的多重 PCR 检测方法,可同时检测山羊粪便中的 G. duodenalis、C. parvum、E. bieneusi 和 Moniezia,适用于快速、大规模筛查绵羊粪便样本中的肠道寄生虫。
{"title":"[Development and preliminary application of a multiplex PCR assay for simultaneous detection of four intestinal parasites in goats].","authors":"Y Li, X Mu, H Xu, X Luo, R Yu, X Xu, L Yang, X Yu, Y Hong","doi":"10.16250/j.32.1374.2024046","DOIUrl":"https://doi.org/10.16250/j.32.1374.2024046","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Objective: &lt;/strong&gt;To develop a multiplex PCR assay for simultaneous detection of four intestinal parasites, including &lt;i&gt;Giardia duodenalis&lt;/i&gt;, &lt;i&gt;Cryptosporidium parvum&lt;/i&gt;, &lt;i&gt;Enterocytozoon bieneusi&lt;/i&gt; and &lt;i&gt;Moniezia&lt;/i&gt;, and to preliminarily evaluate its detection efficiency.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;Four pairs of specific primers were designed based on the conserved sequences of the corresponding genes of &lt;i&gt;G. duodenalis&lt;/i&gt; (GenBank accession number: XM_001710026.2), &lt;i&gt;C. parvum&lt;/i&gt; (GenBank accession number: XM_626998.1), &lt;i&gt;E. bieneusi&lt;/i&gt; (GenBank accession number: KJ719492.1) and &lt;i&gt;Moniezia&lt;/i&gt; (GenBank accession number: OM296991.1) retrieved from the GenBank database, and a multiplex PCR assay for simultaneous detection of &lt;i&gt;G. duodenalis&lt;/i&gt;, &lt;i&gt;C. parvum&lt;/i&gt;, &lt;i&gt;E. bieneusi&lt;/i&gt; and &lt;i&gt;Moniezia&lt;/i&gt; was developed and optimized. A total of 116 fresh goat stool samples were collected from four goat farms in Zhanjiang City, Guangdong Province during the period from October to December 2022, including 96 samples used for evaluating the detection efficacy of the multiplex PCR assay, and 20 samples as baseline controls for sample testing. Genomic DNA extracted from 96 goat stool samples was tested using the single-target PCR assay and the developed multiplex PCR assay, and the sensitivity, specificity, positive predictive value, and negative predictive value of the multiplex PCR assay were evaluated for detection of &lt;i&gt;G. duodenalis&lt;/i&gt;, &lt;i&gt;C. parvum&lt;/i&gt;, &lt;i&gt;E. bieneusi&lt;/i&gt; and &lt;i&gt;Moniezia&lt;/i&gt; DNA in goat stool samples with the single-target PCR assay as the gold standard.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;The multiplex PCR assay developed in this study allowed simultaneous amplification of specific gene fragments of &lt;i&gt;G. duodenalis&lt;/i&gt;, &lt;i&gt;C. parvum&lt;/i&gt;, &lt;i&gt;E. bieneusi&lt;/i&gt; and &lt;i&gt;Moniezia&lt;/i&gt;, with 1 400, 755, 314 bp and 585 bp in sizes, respectively, and the detection limit was 10&lt;sup&gt;2&lt;/sup&gt; and higher copies of parasite DNA clones, while the multiplex PCR assay was negative for gene amplification of &lt;i&gt;Schistosoma japonicum&lt;/i&gt;, &lt;i&gt;Fasciola hepatica&lt;/i&gt;, &lt;i&gt;Echinococcus granulosus&lt;/i&gt;, &lt;i&gt;Blastocystis hominis&lt;/i&gt; and &lt;i&gt;Homalogaster paloniae&lt;/i&gt;. Single-target PCR assay and the developed multiplex PCR assay were employed to test DNA samples extracted from 96 goat stool samples, and single-target PCR assay tested positive in 40 goat stool samples (41.67%), including 39 positive samples tested with the multiplex PCR assay, with a mean coincidence rate of 97.50% (39/40). The multiplex PCR assay tested positive for &lt;i&gt;G. duodenalis&lt;/i&gt; DNA in 26 goat stool samples (27.10%), &lt;i&gt;C. parvum&lt;/i&gt; DNA in 22 samples (22.90%), &lt;i&gt;E. bieneusi&lt;/i&gt; DNA in 24 samples (25.00%), and &lt;i&gt;Moniezia&lt;/i&gt; in 9 samples (9.40%), which was consistent with the detection using the single-target PCR assay. The sensitivity, negative predictive value, and positive predictive value of the multiplex PCR assay were 96.15%, 95.83%, 100.00% and 100.0","PeriodicalId":38874,"journal":{"name":"中国血吸虫病防治杂志","volume":"36 4","pages":"376-383"},"PeriodicalIF":0.0,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142355888","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Auxiliary diagnostic value of T cells spot test of Mycobacterium tuberculosis infection for pulmonary and extra-pulmonary tuberculosis among the elderly]. [结核分枝杆菌感染的 T 细胞斑点试验对老年人肺部和肺外结核病的辅助诊断价值]。
Q3 Medicine Pub Date : 2024-06-18 DOI: 10.16250/j.32.1374.2024121
R Huang, S Li, C Wang

Objective: To evaluate the auxiliary diagnostic value of T cells spot test of Mycobacterium tuberculosis infection (T-SPOT.TB) for pulmonary and extra-pulmonary tuberculosis among the elderly.

Methods: A total of 173 elderly patients at ages of 60 years and older and with suspected tuberculosis that were admitted to People's Hospital of Xinjiang Uygur Autonomous Region during the period from October 2022 through February 2024 were enrolled, and all patients underwent T-SPOT.TB, acid fast staining and GeneXpert MTB/RIF tests. The etiological tests of MTB served as a gold standard, and the diagnostic values of T-SPOT.TB, acid fast staining and GeneXpert MTB/RIF tests for pulmonary and extra-pulmonary tuberculosis were compared among the elderly patients.

Results: Of the 173 elderly patients suspected of tuberculosis, there were 44 patients definitely diagnosed with pulmonary tuberculosis, 30 cases with extra-pulmonary tuberculosis, and 99 cases without tuberculosis. The sensitivities of T-SPOT.TB, acid fast staining and GeneXpert MTB/RIF tests were 86.5%, 27.0% and 54.1% for diagnosis of tuberculosis. The sensitivities of T-SPOT.TB were 86.4% and 86.7% for diagnosis of pulmonary tuberculosis and extra-pulmonary tuberculosis, with an 80.8% specificity for diagnosis of tuberculosis. The sensitivities of GeneXpert MTB/RIF were 56.8% and 50.0% for diagnosis of pulmonary tuberculosis and extra-pulmonary tuberculosis, with a 100.0% specificity each, and the sensitivities of acid fast staining were 31.8% and 20.0% for diagnosis of pulmonary tuberculosis and extra-pulmonary tuberculosis, with a 100.0% specificity each. In addition, the areas under the receiver operating characteristic curve were 0.836, 0.635 and 0.770 for diagnosis of tuberculosis with T-SPOT.TB, acid fast staining and GeneXpert MTB/RIF tests among the elderly patients, respectively.

Conclusions: T-SPOT.TB has a high auxiliary diagnostic value for both pulmonary and extra-pulmonary tuberculosis among elderly patients.

摘要评估结核分枝杆菌感染T细胞点滴试验(T-SPOT.TB)对老年人肺结核和肺外结核的辅助诊断价值:方法:选取2022年10月至2024年2月期间新疆维吾尔自治区人民医院收治的173例60岁及以上疑似肺结核老年患者为研究对象,对所有患者进行T-SPOT.TB、酸性快速染色和GeneXpert MTB/RIF检测。将 MTB 病原学检测作为金标准,比较了 T-SPOT.TB、酸性快速染色和 GeneXpert MTB/RIF 检测对老年患者肺部和肺外结核病的诊断价值:结果:在 173 名疑似肺结核的老年患者中,确诊为肺结核的有 44 例,肺外结核 30 例,无肺结核的 99 例。T-SPOT.TB、酸性快速染色和 GeneXpert MTB/RIF 检测对肺结核诊断的敏感度分别为 86.5%、27.0% 和 54.1%。T-SPOT.TB 对诊断肺结核和肺外结核的敏感性分别为 86.4% 和 86.7%,对诊断结核的特异性为 80.8%。基因Xpert MTB/RIF诊断肺结核和肺外结核的敏感性分别为56.8%和50.0%,特异性均为100.0%;酸性快速染色诊断肺结核和肺外结核的敏感性分别为31.8%和20.0%,特异性均为100.0%。此外,用 T-SPOT.TB、酸性快速染色法和 GeneXpert MTB/RIF 检测诊断老年患者肺结核的接收者操作特征曲线下面积分别为 0.836、0.635 和 0.770:结论:T-SPOT.TB 对老年患者的肺部和肺外结核病都有很高的辅助诊断价值。
{"title":"[Auxiliary diagnostic value of T cells spot test of <i>Mycobacterium tuberculosis</i> infection for pulmonary and extra-pulmonary tuberculosis among the elderly].","authors":"R Huang, S Li, C Wang","doi":"10.16250/j.32.1374.2024121","DOIUrl":"https://doi.org/10.16250/j.32.1374.2024121","url":null,"abstract":"<p><strong>Objective: </strong>To evaluate the auxiliary diagnostic value of T cells spot test of <i>Mycobacterium tuberculosis</i> infection (T-SPOT.TB) for pulmonary and extra-pulmonary tuberculosis among the elderly.</p><p><strong>Methods: </strong>A total of 173 elderly patients at ages of 60 years and older and with suspected tuberculosis that were admitted to People's Hospital of Xinjiang Uygur Autonomous Region during the period from October 2022 through February 2024 were enrolled, and all patients underwent T-SPOT.TB, acid fast staining and GeneXpert MTB/RIF tests. The etiological tests of MTB served as a gold standard, and the diagnostic values of T-SPOT.TB, acid fast staining and GeneXpert MTB/RIF tests for pulmonary and extra-pulmonary tuberculosis were compared among the elderly patients.</p><p><strong>Results: </strong>Of the 173 elderly patients suspected of tuberculosis, there were 44 patients definitely diagnosed with pulmonary tuberculosis, 30 cases with extra-pulmonary tuberculosis, and 99 cases without tuberculosis. The sensitivities of T-SPOT.TB, acid fast staining and GeneXpert MTB/RIF tests were 86.5%, 27.0% and 54.1% for diagnosis of tuberculosis. The sensitivities of T-SPOT.TB were 86.4% and 86.7% for diagnosis of pulmonary tuberculosis and extra-pulmonary tuberculosis, with an 80.8% specificity for diagnosis of tuberculosis. The sensitivities of GeneXpert MTB/RIF were 56.8% and 50.0% for diagnosis of pulmonary tuberculosis and extra-pulmonary tuberculosis, with a 100.0% specificity each, and the sensitivities of acid fast staining were 31.8% and 20.0% for diagnosis of pulmonary tuberculosis and extra-pulmonary tuberculosis, with a 100.0% specificity each. In addition, the areas under the receiver operating characteristic curve were 0.836, 0.635 and 0.770 for diagnosis of tuberculosis with T-SPOT.TB, acid fast staining and GeneXpert MTB/RIF tests among the elderly patients, respectively.</p><p><strong>Conclusions: </strong>T-SPOT.TB has a high auxiliary diagnostic value for both pulmonary and extra-pulmonary tuberculosis among elderly patients.</p>","PeriodicalId":38874,"journal":{"name":"Chinese Journal of Schistosomiasis Control","volume":"36 3","pages":"310-313"},"PeriodicalIF":0.0,"publicationDate":"2024-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141477575","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Molecular tracing of Biomphalaria straminea in China]. [中国石斑鱼的分子追踪]。
Q3 Medicine Pub Date : 2024-06-18 DOI: 10.16250/j.32.1374.2024069
L Duan, L Qu, Y Guo, W Gu, S Lü, Y Zhang, X Zhou

Objective: To investigate the origin of Biomphalaria straminea in China, so as to provide insights into assessment of schistosomiasis mansoni transmission risk and B. straminea control.

Methods: Guanlan River, Dasha River, Shenzhen Reservoir, upper and lower reaches of Kuiyong River, and Xinzhen River in Shenzhen, China, were selected as sampling sites. Ten Biomphalaria samples were collected from each site, and genomic DNA was extracted from Biomphalaria samples. DNA samples were obtained from 15 B. straminea sampled from 5 sampling sites in Minas Gerais State, Pará State, Federal District, Pernambuco State, and Sao Paulo State in Brazil, South America. Cytochrome c oxidase I (COI) and mitochondrial 16S ribosomal RNA (16S rRNA) genes were sampled using the above DNA templates, and the amplified products were sequenced. The COI and 16S rRNA gene sequences were downloaded from GenBank, and the sampling sites were acquired. All COI and 16S rRNA gene sequences were aligned and evolutionary trees of B. straminea were created based on COI and 16S rRNA gene sequences to identify the genetic similarity and evolutionary relationship between B. straminea samples from China and South America.

Results: A total of 60 COI gene sequences with a length of 529 bp and 3 haplotypes were obtained from B. straminea sampled from China. There were 165 COI gene sequences of B. straminea retrieved from GenBank, and following alignment with the above 60 gene sequences, a total of 33 haplotypes were obtained. Phylogenetic analysis showed that the three haplotypes of B. straminea from China were clustered into one clade, among which the haplotype China11 and three B. straminea samples from Brazil retrieved from GenBank belonged to the same haplotype. Geographical evolution analysis showed that the B. straminea samples from three sampling sites along eastern coasts of Brazil had the same haplotype with China11, and B. straminea samples from other two sampling sites were closely, genetically related to China11. A total of 60 16S rDNA gene sequences with approximately 322 bp in length were amplified from B. straminea in China, with 2 haplotypes identified. A total of 70 16S rDNA gene sequences of B. straminea were captured from GenBank. Phylogenetic analysis showed that Biomphalaria snails collected from China were clustered into a clade, and the haplotype China64 and the haplotype 229BS from Brazil shared the same haplotype. The 49 16S rDNA gene sequences of B. straminea from 25 sampling sites in southern Brazil, which were captured from GenBank, were included in the present analysis, and the B. straminea from 3 sampling sites shared the same haplotype with China64 in China. Geographical evolution

目的调查中国疟原虫的来源,为评估曼氏血吸虫病传播风险和疟原虫防治提供依据:方法:选取中国深圳市观澜河、大沙河、深圳水库、魁永河上下游和新圳河为采样点。在每个采样点采集了 10 个画眉藻样本,并从画眉藻样本中提取了基因组 DNA。从南美洲巴西米纳斯吉拉斯州、帕拉州、联邦区、伯南布哥州和圣保罗州的 5 个采样点采集了 15 个 B. straminea 样本,并从中提取了基因组 DNA。利用上述 DNA 模板对细胞色素 c 氧化酶 I(COI)和线粒体 16S 核糖体 RNA(16S rRNA)基因进行了采样,并对扩增产物进行了测序。从 GenBank 下载了 COI 和 16S rRNA 基因序列,并获取了取样位点。对所有 COI 和 16S rRNA 基因序列进行比对,并根据 COI 和 16S rRNA 基因序列建立 B. straminea 的进化树,以确定中国和南美洲 B. straminea 样本之间的遗传相似性和进化关系:结果:共获得了 60 个 COI 基因序列,长度为 529 bp,3 个单倍型。从 GenBank 中检索到 165 条 B. straminea 的 COI 基因序列,与上述 60 条基因序列比对后,共得到 33 个单倍型。系统进化分析表明,来自中国的三个单倍型被聚为一个支系,其中单倍型 China11 与从 GenBank 中检索到的三个来自巴西的 B. straminea 样品属于同一单倍型。地理进化分析表明,巴西东部沿海三个采样点的 B. straminea 样品与 China11 具有相同的单倍型,另外两个采样点的 B. straminea 样品与 China11 具有密切的遗传亲缘关系。从中国的 B. straminea 共扩增出 60 个长度约为 322 bp 的 16S rDNA 基因序列,并确定了 2 个单倍型。从 GenBank 中获取了 70 个 B. straminea 的 16S rDNA 基因序列。系统进化分析表明,从中国采集的 Bomphalaria 蜗牛聚为一个支系,单倍型 China64 与来自巴西的单倍型 229BS 具有相同的单倍型。从 GenBank 中获取的巴西南部 25 个采样点的 49 个 B. straminea 的 16S rDNA 基因序列被纳入本分析,其中 3 个采样点的 B. straminea 与中国的 China64 具有相同的单倍型。基于 COI 和 16S rRNA 基因序列的地理进化分析表明,巴西东部沿海地区采样的 B. straminea 与中国采集的 Biomphalaria 螺类在两个基因片段序列上具有相同的单倍型:结论:中国田螺的特点是遗传多样性较低。中国田螺与巴西东部沿海地区田螺的遗传相似性较高,可能起源于巴西东部沿海地区。
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引用次数: 0
[Application of the CRISPR/Cas system in gene editing and nucleic acid detection of parasitic diseases: a review]. [CRISPR/Cas 系统在寄生虫病基因编辑和核酸检测中的应用:综述]。
Q3 Medicine Pub Date : 2024-06-17 DOI: 10.16250/j.32.1374.2024057
S Yan, S Yang, H Yang, Y Xin, B Xu, W Hu, Y Lu, B Zheng

CRISPR/Cas system, an adaptive immune system with clustered regularly interspaced short palindromic repeats, may interfere with exogenous nucleic acids and protect prokaryotes from external damages, is an effective gene editing and nucleic acid detection tools. The CRISPR/Cas system has been widely applied in virology and bacteriology; however, there is relatively less knowledge about the application of the CRISPR/Cas system in parasitic diseases. The review summarizes the mechanisms of action of the CRISPR/Cas system and provides a comprehensive overview of their application in gene editing and nucleic acid detection of parasitic diseases, so as to provide insights into future studies on parasitic diseases.

CRISPR/Cas系统是一种具有簇状规则间隔短回文重复序列的适应性免疫系统,可干扰外源核酸,保护原核生物免受外界损害,是一种有效的基因编辑和核酸检测工具。CRISPR/Cas系统已广泛应用于病毒学和细菌学领域,但在寄生虫病领域的应用却相对较少。本综述总结了CRISPR/Cas系统的作用机制,并全面介绍了其在寄生虫病基因编辑和核酸检测中的应用,以期为今后的寄生虫病研究提供启示。
{"title":"[Application of the CRISPR/Cas system in gene editing and nucleic acid detection of parasitic diseases: a review].","authors":"S Yan, S Yang, H Yang, Y Xin, B Xu, W Hu, Y Lu, B Zheng","doi":"10.16250/j.32.1374.2024057","DOIUrl":"https://doi.org/10.16250/j.32.1374.2024057","url":null,"abstract":"<p><p>CRISPR/Cas system, an adaptive immune system with clustered regularly interspaced short palindromic repeats, may interfere with exogenous nucleic acids and protect prokaryotes from external damages, is an effective gene editing and nucleic acid detection tools. The CRISPR/Cas system has been widely applied in virology and bacteriology; however, there is relatively less knowledge about the application of the CRISPR/Cas system in parasitic diseases. The review summarizes the mechanisms of action of the CRISPR/Cas system and provides a comprehensive overview of their application in gene editing and nucleic acid detection of parasitic diseases, so as to provide insights into future studies on parasitic diseases.</p>","PeriodicalId":38874,"journal":{"name":"Chinese Journal of Schistosomiasis Control","volume":"36 3","pages":"314-320"},"PeriodicalIF":0.0,"publicationDate":"2024-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141477643","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Effect of oxymatrine on Cryptosporidium parvum infection in mice based on the HMGB1-TLR2/TLR4-NF-κB pathway]. [基于 HMGB1-TLR2/TLR4-NF-κB 途径的氧化苦参碱对小鼠感染副隐孢子虫的影响]。
Q3 Medicine Pub Date : 2024-06-17 DOI: 10.16250/j.32.1374.2024019
J Shi, R Ji, Z Guan, X Zhang, Y Lu

Objective: To investigate the involvement of the high mobility group box protein B1 (HMGB1)-Toll-like receptor 2 (TLR2)/TLR4-nuclear factor κB (NF-κB) pathway in the intestinal mucosal injury induced by Cryptosporidium parvum infection, and to examine the effect of oxymatrine (OMT) on C. parvum infection in mice.

Methods: Forty SPF 4-week-old BALB/c mice were randomly divided into four groups, including the control group, infection group, glycyrrhizin (GA) group and OMT group. Each mouse was orally administered with 1 × 105 C. parvum oocysts one week in the infection, GA and OMT groups following dexamethasone-induced immunosuppression to model C. parvum intestinal infections in mice. Upon successful modeling, mice in the GA group were intraperitoneally injected with GA at a daily dose of 25.9 mL/kg for successive two weeks, and animals in the OMT group were orally administered OMT at a daily dose of 50 mg/kg for successive two weeks, while mice in the control group were given normal food and water. All mice were sacrificed two weeks post-treatment, and proximal jejunal tissues were sampled. The pathological changes of mouse intestinal mucosal specimens were observed using hematoxylin-eosin (HE) staining, and the mouse intestinal villous height, intestinal crypt depth and the ratio of intestinal villous height to intestinal crypt depth were measured. The occludin and zonula occludens protein 1 (ZO1) expression was determined in mouse intestinal epithelial cells using immunohistochemistry, and the relative expression of HMGB1, TLR2, TLR4, myeloid differentiation primary response gene 88 (MyD88) and NF-κB p65 mRNA was quantified in mouse jejunal tissues using quantitative real-time PCR (qPCR) assay.

Results: HE staining showed that the mouse intestinal villi were obviously atrophic, shortened, and detached, and the submucosal layer of the mouse intestine was edematous in the infection group as compared with the control group, while the mouse intestinal villi tended to be structurally intact and neatly arranged in the GA and OMT groups. There were significant differences among the four groups in terms of the mouse intestinal villous height (F = 6.207, P = 0.000 5), intestinal crypt depth (F = 6.903, P = 0.000 3) and the ratio of intestinal villous height to intestinal crypt depth (F = 37.190, P < 0.000 1). The mouse intestinal villous height was lower in the infection group than in the control group [(321.9 ± 41.1) μm vs. (399.5 ± 30.9) μm; t = 4.178, P < 0.01] and the GA group [(321.9 ± 41.1) μm vs. (383.7 ± 42.7) μm; t = 3.130, P < 0.01], and the mouse intestinal crypt depth was greater in the infection group [(185.0 ± 35.9) μm] than in the control group [(128.4 ± 23.6) μm] (t = 3.877, P < 0.01) and GA group [(143.3 ± 24.7) μm]

目的研究高迁移率基团盒蛋白B1(HMGB1)-类托尔受体2(TLR2)/TLR4-核因子κB(NF-κB)通路参与副猪隐孢子虫感染引起的肠黏膜损伤,并探讨氧化苦参碱(OMT)对小鼠副猪隐孢子虫感染的影响:方法:将 40 只 SPF 4 周龄 BALB/c 小鼠随机分为四组,包括对照组、感染组、甘草酸苷(GA)组和 OMT 组。感染组、甘草酸苷组和 OMT 组的每只小鼠在地塞米松诱导的免疫抑制后一周口服 1 × 105 个副猪嗜血杆菌卵囊,以建立副猪嗜血杆菌肠道感染模型。成功建模后,GA 组小鼠腹腔注射 GA,每天剂量为 25.9 mL/kg,连续两周;OMT 组小鼠口服 OMT,每天剂量为 50 mg/kg,连续两周;对照组小鼠给予正常食物和水。所有小鼠在治疗后两周处死,并采集空肠近端组织样本。用苏木精-伊红(HE)染色法观察小鼠肠粘膜标本的病理变化,并测量小鼠肠绒毛高度、肠隐窝深度以及肠绒毛高度与肠隐窝深度的比值。采用免疫组化方法测定了小鼠肠上皮细胞中的闭锁素和闭锁带蛋白1(ZO1)的表达,并采用实时定量PCR(qPCR)方法测定了小鼠空肠组织中HMGB1、TLR2、TLR4、髓样分化主要反应基因88(MyD88)和NF-κB p65 mRNA的相对表达量:HE染色显示:与对照组相比,感染组小鼠肠绒毛明显萎缩、缩短、脱落,肠黏膜下层水肿;而GA组和OMT组小鼠肠绒毛结构完整,排列整齐。四组小鼠肠绒毛高度(F = 6.207,P = 0.000 5)、肠隐窝深度(F = 6.903,P = 0.000 3)和肠绒毛高度与肠隐窝深度之比(F = 37.190,P < 0.000 1)均有显著差异。感染组小鼠肠绒毛高度低于对照组[(321.9 ± 41.1)μm vs. (399.5 ± 30.9)μm;t = 4.178,P < 0.01],也低于 GA 组[(321.9 ± 41.1)μm vs. (383.7 ± 42.7)μm;t = 3.感染组小鼠肠隐窝深度[(185.0 ± 35.9)μm]大于对照组[(128.4 ± 23.6)μm](t = 3.877,P < 0.01)和 GA 组[(143.3 ± 24.7)μm](t = 2.710,P < 0.05)。OMT组小鼠肠绒毛高度[(375.3 ± 22.9)μm]大于感染组(t = 3.888,P < 0.01),OMT组与对照组小鼠肠绒毛高度无显著差异(t = 1.989,P > 0.05)。OMT组小鼠肠隐窝深度[(121.5 ± 27.3)μm]明显低于感染组(t = 4.133,P < 0.01),OMT组与对照组小鼠肠隐窝深度无明显差异(t = 0.575,P > 0.05)。感染组小鼠肠绒毛高度与肠隐窝深度之比(1.8 ± 0.2)显著低于对照组(3.1 ± 0.3)(t = 10.540,P < 0.01)和 GA 组(2.7 ± 0.3)(t = 7.370,P < 0.01),OMT 组小鼠肠绒毛高度与肠隐窝深度之比(3.1 ± 0.2)明显高于感染组(t = 15.020,P < 0.01);但OMT组与对照组小鼠肠绒毛高度与肠隐窝深度之比无明显差异(t = 0.404,P > 0.05)。免疫组化染色显示,四组小鼠肠上皮细胞的闭塞素(F = 28.031,P < 0.000 1)和ZO1表达(F = 14.122,P < 0.000 1)存在显著差异。感染组小鼠肠上皮细胞中阳性闭锁表达的比例明显低于对照组[(14.3 ± 4.5)% vs. (28.3 ± 0.5)%;t = 3.810,P < 0.01],GA 组小鼠肠上皮细胞中阳性闭锁表达的比例明显高于对照组[(30.3±1.3)%]和 OMT 组[(25.8±1.5)%]明显高于感染组(t = 7.620 和 5.391,P 值均<0.01);但 GA 组或 OMT 组与对照组小鼠肠上皮细胞的阳性闭塞表达比例无明显差异(t = 1.791 和 2.033,P 值均>0.05)。
{"title":"[Effect of oxymatrine on <i>Cryptosporidium parvum</i> infection in mice based on the HMGB1-TLR2/TLR4-NF-κB pathway].","authors":"J Shi, R Ji, Z Guan, X Zhang, Y Lu","doi":"10.16250/j.32.1374.2024019","DOIUrl":"https://doi.org/10.16250/j.32.1374.2024019","url":null,"abstract":"<p><strong>Objective: </strong>To investigate the involvement of the high mobility group box protein B1 (HMGB1)-Toll-like receptor 2 (TLR2)/TLR4-nuclear factor κB (NF-κB) pathway in the intestinal mucosal injury induced by <i>Cryptosporidium parvum</i> infection, and to examine the effect of oxymatrine (OMT) on <i>C. parvum infection</i> in mice.</p><p><strong>Methods: </strong>Forty SPF 4-week-old BALB/c mice were randomly divided into four groups, including the control group, infection group, glycyrrhizin (GA) group and OMT group. Each mouse was orally administered with 1 × 10<sup>5</sup> <i>C. parvum</i> oocysts one week in the infection, GA and OMT groups following dexamethasone-induced immunosuppression to model <i>C. parvum</i> intestinal infections in mice. Upon successful modeling, mice in the GA group were intraperitoneally injected with GA at a daily dose of 25.9 mL/kg for successive two weeks, and animals in the OMT group were orally administered OMT at a daily dose of 50 mg/kg for successive two weeks, while mice in the control group were given normal food and water. All mice were sacrificed two weeks post-treatment, and proximal jejunal tissues were sampled. The pathological changes of mouse intestinal mucosal specimens were observed using hematoxylin-eosin (HE) staining, and the mouse intestinal villous height, intestinal crypt depth and the ratio of intestinal villous height to intestinal crypt depth were measured. The occludin and zonula occludens protein 1 (ZO1) expression was determined in mouse intestinal epithelial cells using immunohistochemistry, and the relative expression of <i>HMGB1</i>, <i>TLR2</i>, <i>TLR4</i>, myeloid differentiation primary response gene 88 (<i>MyD88</i>) and <i>NF-κB p65 mRNA</i> was quantified in mouse jejunal tissues using quantitative real-time PCR (qPCR) assay.</p><p><strong>Results: </strong>HE staining showed that the mouse intestinal villi were obviously atrophic, shortened, and detached, and the submucosal layer of the mouse intestine was edematous in the infection group as compared with the control group, while the mouse intestinal villi tended to be structurally intact and neatly arranged in the GA and OMT groups. There were significant differences among the four groups in terms of the mouse intestinal villous height (<i>F</i> = 6.207, <i>P</i> = 0.000 5), intestinal crypt depth (<i>F</i> = 6.903, <i>P</i> = 0.000 3) and the ratio of intestinal villous height to intestinal crypt depth (<i>F</i> = 37.190, <i>P</i> < 0.000 1). The mouse intestinal villous height was lower in the infection group than in the control group [(321.9 ± 41.1) μm vs. (399.5 ± 30.9) μm; <i>t</i> = 4.178, <i>P</i> < 0.01] and the GA group [(321.9 ± 41.1) μm vs. (383.7 ± 42.7) μm; <i>t</i> = 3.130, <i>P</i> < 0.01], and the mouse intestinal crypt depth was greater in the infection group [(185.0 ± 35.9) μm] than in the control group [(128.4 ± 23.6) μm] (<i>t</i> = 3.877, <i>P</i> < 0.01) and GA group [(143.3 ± 24.7) μm] ","PeriodicalId":38874,"journal":{"name":"Chinese Journal of Schistosomiasis Control","volume":"36 3","pages":"286-293"},"PeriodicalIF":0.0,"publicationDate":"2024-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141477582","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
期刊
中国血吸虫病防治杂志
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