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Analysis of the Class 1 Integrons, Carbapenemase Genes and Biofilm Formation Genes Occurrence in Acinetobacter baumannii Clinical Isolates. 鲍曼不动杆菌临床分离株中 1 类整合子、碳青霉烯酶基因和生物膜形成基因的发生率分析。
Pub Date : 2024-05-29 eCollection Date: 2024-06-01 DOI: 10.33073/pjm-2024-017
Yu Xiu, Yueru Dai, Shasha Yin, Quhao Wei

Acinetobacter baumannii is a non-fermentative Gram-negative bacterium that can cause nosocomial infections in critically ill patients. Carbapenem-resistant A. baumannii (CRAB) has spread rapidly in clinical settings and has become a key concern. The main objective of this study was to identify the distribution of integrons and biofilm-formation-related virulence genes in CRAB isolates. A total of 269 A. baumannii isolates (219 isolates of CRAB and 50 isolates of carbapenem-sensitive A. baumannii (CSAB)) were collected. Carbapenemase genes (bla KPC, bla VIM, bla IMP, bla NDM, and bla OXA-23-like) and biofilm-formation-related virulence genes (abal, bfms, bap, and cusE) were screened with PCR. Class 1 integron was screened with PCR, and common promoters and gene cassette arrays were determined with restriction pattern analysis combined with primer walking sequencing. Whole-genome sequencing was conducted, and data were analyzed for a bla OXA-23-like-negative isolate. All 219 CRAB isolates were negative for bla KPC, bla VIM, bla IMP, and bla NDM, while bla OXA-23-like was detected in 218 isolates. The detection rates for abal, bfms, bap, and cusE in 219 CRAB were 93.15%, 63.93%, 88.13%, and 77.63%, respectively. Class 1 integron was detected in 75 CRAB (34.25%) and in 3 CSAB. The single gene cassette array aacA4-catB8-aadA1 with relatively strong PcH2 promoter was detected in class 1 integrons. The bla OXA-23-like-negative CRAB isolate was revealed to be a new sequence type (Oxford 3272, Pasteur 2520) carrying bla OXA-72, bla OXA-259, and bla ADC-26. In conclusion, bla OXA-23-like was the main reason for CRAB's resistance to carbapenems. A new (Oxford 3272, Pasteur 2520) CRAB sequence type carrying the bla OXA-72, bla OXA-259, and bla ADC-26 was reported.

鲍曼不动杆菌(Acinetobacter baumannii)是一种非发酵革兰氏阴性细菌,可引起重症患者的院内感染。耐碳青霉烯鲍曼不动杆菌(CRAB)已在临床环境中迅速传播,并已成为人们关注的焦点。本研究的主要目的是鉴定 CRAB 分离物中整合子和生物膜形成相关毒力基因的分布。本研究共收集了269株鲍曼不动杆菌分离株(219株CRAB分离株和50株碳青霉烯类敏感鲍曼不动杆菌(CSAB)分离株)。用 PCR 筛选了碳青霉烯酶基因(bla KPC、bla VIM、bla IMP、bla NDM 和 bla OXA-23-like)和与生物膜形成相关的毒力基因(abal、bfms、bap 和 cusE)。用 PCR 筛选了 1 类整合子,并通过限制性模式分析和引物走行测序确定了常见启动子和基因盒阵列。进行了全基因组测序,并对 bla OXA-23 样阴性分离物进行了数据分析。所有 219 个 CRAB 分离物的 bla KPC、bla VIM、bla IMP 和 bla NDM 均为阴性,而 218 个分离物中检测到了 bla OXA-23-like。在 219 个 CRAB 分离物中,abal、bfms、bap 和 cusE 的检出率分别为 93.15%、63.93%、88.13% 和 77.63%。在 75 个 CRAB(34.25%)和 3 个 CSAB 中检测到了 1 类整合子。在 1 类整合子中检测到了带有相对较强的 PcH2 启动子的单基因盒阵列 aacA4-catB8-aadA1。bla OXA-23 样阴性的 CRAB 分离物被发现是一种新的序列类型(Oxford 3272,Pasteur 2520),携带 bla OXA-72、bla OXA-259 和 bla ADC-26。总之,类 bla OXA-23 是 CRAB 对碳青霉烯类产生耐药性的主要原因。报告了一种携带 bla OXA-72、bla OXA-259 和 bla ADC-26 的新 CRAB 序列类型(牛津 3272,巴斯德 2520)。
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引用次数: 0
Epidemiological Characteristics of Shiga Toxin-Producing Escherichia coli Responsible for Infections in the Polish Pediatric Population. 导致波兰儿科人群感染的产志贺毒素大肠杆菌的流行病学特征。
Pub Date : 2024-05-10 eCollection Date: 2024-06-01 DOI: 10.33073/pjm-2024-016
Dominika Seliga-Gąsior, Beata Sokól-Leszczyñska, Jolanta Krzysztoñ-Russjan, Diana Wierzbicka, Karolina Stępieñ-Hołubczat, Paulina Lewandowska, Ewa Frankiewicz, Andrzej Cacko, Beata Leszczyñska, Urszula Demkow, Edyta Podsiadły

Shiga toxin-producing Escherichia coli (STEC) are zoonotic pathogens causing hemorrhagic colitis and hemolytic uremic syndrome (HUS) in children and the elderly. Stool samples were collected from 180 children hospitalized in five pediatric centers in Poland in 2018-2022. Direct stx1/stx2 gene detection by PCR in feces and E. coli isolates was performed. Antibiotic susceptibility was tested according to EUCAST v.12. Randomly selected isolates were serotyped with O157 antiserum and genotyped by pulsed-field gel electrophoresis (PFGE). A total of 44 E. coli isolates were confirmed as STEC by PCR. Among them, 84.4% were positive for stx2, and equally 6,8% for only stx1 and both stx1 and stx2 genes. The stx1 gene was also found in one Citrobacter freundii isolate. E. coli serotype O157 was present in 97.6% of the isolates. STEC infections most often occurred between June-October with a peak in July and August (51%). The highest, 77.8% of STEC isolates were found in the 1-5 years old group. No extended-spectrum β-lactamases (ESBL) were found. Resistance only to amoxicillin/clavulanic acid (24.4%), piperacillin/tazobactam (3%), cefotaxime (6%), gentamicin (6%), ciprofloxacin (3%), azithromycin (3%), trimethoprim/sulfamethoxazole (24,2%) was detected. PFGE analysis showed 18 PFGE types with no clonal distribution. Eight isolates with A, B, and C PFGE types showed genetic relatedness in the type with no detection of transmission way of distribution. STEC strains pose a serious threat to human health, therefore demographic and epidemiological characteristics are crucial for their surveillance.

产志贺毒素大肠杆菌(STEC)是人畜共患的病原体,可导致儿童和老年人出血性结肠炎和溶血性尿毒症(HUS)。2018-2022年,波兰5家儿科中心收集了180名住院儿童的粪便样本。通过 PCR 直接检测粪便和大肠杆菌分离物中的 stx1/stx2 基因。抗生素敏感性根据 EUCAST v.12 进行检测。用 O157 抗血清对随机挑选的分离物进行血清分型,并通过脉冲场凝胶电泳(PFGE)进行基因分型。通过聚合酶链式反应(PCR),共确认了 44 个大肠杆菌分离物为 STEC。其中,84.4%对stx2基因呈阳性,6.8%仅对stx1基因呈阳性,6.8%同时对stx1和stx2基因呈阳性。在一个弗氏柠檬杆菌分离物中也发现了 stx1 基因。97.6%的分离物中含有大肠杆菌血清型 O157。STEC 感染最常发生在 6 月至 10 月间,7 月和 8 月为高峰期(51%)。1-5 岁年龄组的 STEC 感染率最高,占 77.8%。没有发现扩谱β-内酰胺酶(ESBL)。仅检测出对阿莫西林/克拉维酸(24.4%)、哌拉西林/他唑巴坦(3%)、头孢他啶(6%)、庆大霉素(6%)、环丙沙星(3%)、阿奇霉素(3%)、三甲双胍/磺胺甲噁唑(24.2%)的耐药性。PFGE 分析显示有 18 种 PFGE 类型,无克隆分布。8株具有A、B和C PFGE类型的分离株显示出类型中的遗传相关性,但未检测到传播分布方式。STEC 菌株对人类健康构成严重威胁,因此人口统计学和流行病学特征对其监测至关重要。
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引用次数: 0
Investigation of Azole Resistance Involving cyp51A and cyp51B Genes in Clinical Aspergillus flavus Isolates. 临床黄曲霉菌分离物中涉及 cyp51A 和 cyp51B 基因的唑类抗性研究
Pub Date : 2024-05-03 eCollection Date: 2024-06-01 DOI: 10.33073/pjm-2024-001
Dhoha Ghorbel, Imen Amouri, Nahed Khemekhem, Sourour Neji, Houaida Trabelsi, Moez Elloumi, Hayet Sellami, Fattouma Makni, Ali Ayadi, Ines Hadrich

This study aimed to investigate azole resistance mechanisms in Aspergillus flavus, which involve cyp51A and cyp51B genes. Real-time Reverse Transcriptase qPCR method was applied to determine the overexpression of cyp51A and cyp51B genes for 34 A. flavus isolates. PCR sequencing of these two genes was used to detect the presence of gene mutations. Susceptibility test found sensitivity to voriconazole (VOR) in all strains. 14.7% and 8.8% of isolates were resistant to itraconazole (IT) and posaconazole (POS), respectively, with a cross-resistance in 5.8%. For the double resistant isolates (IT/POS), the expression of cyp51A was up to 17-fold higher. PCR sequencing showed the presence of 2 mutations in cyp51A: a synonymous point mutation (P61P) in eight isolates, which did not affect the structure of CYP51A protein, and another non synonymous mutation (G206L) for only the TN-33 strain (cross IT/POS resistance) causing an amino acid change in the protein sequence. However, we noted in cyp51B the presence of the only non-synonymous mutation (L177G) causing a change in amino acids in the protein sequence for the TN-31 strain, which exhibits IT/POS cross-resistance. A short single intron of 67 bp was identified in the cyp51A gene, whereas three short introns of 54, 53, and 160 bp were identified in the cyp51B gene. According to the models provided by PatchDock software, the presence of non-synonymous mutations did not affect the interaction of CYP51A and CYP51B proteins with antifungals. In our study, the overexpression of the cyp51A and cyp51B genes is the primary mechanism responsible for resistance in A. flavus collection. Nevertheless, other resistance mechanisms can be involved.

本研究旨在探讨黄曲霉的唑类抗性机制,其中涉及cyp51A和cyp51B基因。采用实时逆转录酶 qPCR 方法测定了 34 株黄曲霉分离株的 cyp51A 和 cyp51B 基因的过表达情况。通过对这两个基因进行 PCR 测序,检测是否存在基因突变。药敏试验发现,所有菌株都对伏立康唑(VOR)敏感。14.7%和8.8%的分离株分别对伊曲康唑(IT)和泊沙康唑(POS)产生耐药性,其中5.8%产生交叉耐药性。对于双重耐药的分离物(IT/POS),cyp51A的表达量最多高出17倍。PCR 测序显示 cyp51A 存在 2 个突变:8 个分离株存在同义点突变(P61P),但不影响 CYP51A 蛋白的结构;另一个非同义突变(G206L)仅存在于 TN-33 株系(IT/POS 交叉抗性)中,导致蛋白质序列发生氨基酸变化。然而,我们注意到,在 Cyp51B 中,唯一的非同义突变(L177G)导致 TN-31 株蛋白序列中氨基酸的变化,而 TN-31 株表现出 IT/POS 交叉抗性。在 cyp51A 基因中发现了一个 67 bp 的短内含子,而在 cyp51B 基因中发现了三个分别为 54、53 和 160 bp 的短内含子。根据 PatchDock 软件提供的模型,非同义突变的存在并不影响 CYP51A 和 CYP51B 蛋白与抗真菌药物的相互作用。在我们的研究中,cyp51A 和 cyp51B 基因的过度表达是导致黄曲霉产生抗药性的主要机制。不过,其他抗性机制也可能参与其中。
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引用次数: 0
Short-Chain Carboxylates Facilitate the Counting of Yeasts in Sub-High Temperature Daqu. 短链羧酸有助于亚高温大曲中酵母菌的计数
Pub Date : 2024-04-28 eCollection Date: 2024-06-01 DOI: 10.33073/pjm-2024-015
Zhiqiang Ren, Juan Xie, Tuoxian Tang, Zhiguo Huang

Sub-high temperature Daqu, a traditional solid fermenting agent used in Chinese strong-aroma Baijiu production, is abundant in diverse microorganisms, including bacteria, yeasts, molds, and actinomycetes. Among these, yeasts are pivotal for ethanol production and flavor formation. However, counting yeasts in Daqu is challenging due to interference from molds and bacteria. Antibiotics are employed to inhibit bacterial growth, but there is no practical way to suppress molds without affecting the growth of yeasts. In this study, short-chain carboxylates (C1-C6) were added to the culture medium at various pH conditions to investigate their effects on the growth of molds and yeasts. The results demonstrated distinct inhibitory effects of the short-chain carboxylates, depending on both pH and concentration. Several tested short-chain carboxylates effectively suppressed mold growth on agar plates while leaving yeast growth unaffected. This suggests a simple and feasible method for enhancing the efficiency of yeast isolation and counting in Daqu. Such an approach is valuable for studying yeasts in diverse and complex habitats.

亚高温大曲是中国浓香型白酒生产中使用的一种传统固体发酵剂,富含多种微生物,包括细菌、酵母菌、霉菌和放线菌。其中,酵母菌对乙醇的生产和风味的形成至关重要。然而,由于受到霉菌和细菌的干扰,大曲中酵母菌的计数具有挑战性。人们使用抗生素来抑制细菌的生长,但没有切实可行的方法在不影响酵母生长的情况下抑制霉菌的生长。本研究将短链羧酸盐(C1-C6)添加到不同 pH 值条件下的培养基中,以研究它们对霉菌和酵母菌生长的影响。结果表明,短链羧酸盐的抑制作用因 pH 值和浓度而异。几种测试的短链羧酸盐有效抑制了琼脂平板上霉菌的生长,而酵母菌的生长不受影响。这为提高大曲中酵母菌的分离和计数效率提供了一种简单可行的方法。这种方法对于研究多种复杂生境中的酵母菌很有价值。
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引用次数: 0
Purification and Characterization of a Small Thermostable Protease from Streptomyces sp. CNXK100. 链霉菌 CNXK100 小型恒温蛋白酶的纯化与特性分析
Pub Date : 2024-04-28 eCollection Date: 2024-06-01 DOI: 10.33073/pjm-2024-014
Tan Viet Pham, Truong Chinh Hua, Ngoc An Nguyen, Hanh Thi Dieu Nguyen

Proteases derived from Streptomyces demonstrate numerous commendable properties, rendering it extensively applicable in biotechnology and various industrial sectors. This study focused on the purification and characterization of the thermostable protease obtained from Streptomyces sp. CNXK100. The purified protease exhibited an estimated molecular weight of 27 kDa, with optimal activity at 75°C and pH 8.0. Notably, the enzyme remained active even without any metal ions and fully active in the presence of Na+, K+, Mg2+, and Cu2+metal ions. The kinetic parameters were determined with a KM value of 3.13 mg/ml and a Vmax value of 3.28 × 106 U/mg. Furthermore, the protease has demonstrated notable stability when subjected to a treatment temperature of up to 65°C for 60 minutes, and across a broad pH range extending from 5.0 to 10.0. This protease also demonstrated resilience against a spectrum of harsh conditions, including exposure to organic solvents, surfactants, bleaching agents, and proteolytic enzymes. Additionally, the enzyme maintained its activity following treatment with commercial detergents, accomplishing complete thrombus lysis at a concentration of 2.50 mg/ml within 4 hours. Remarkably, the protease exhibited stability in terms of activity and protein concentration for 70 days at 4°C. These findings underscore the potential industrial applications of the thermostable protease from Streptomyces sp. CNXK100.

从链霉菌中提取的蛋白酶具有许多值得称道的特性,因此可广泛应用于生物技术和各种工业领域。本研究的重点是纯化和鉴定从链霉菌 CNXK100 中获得的恒温蛋白酶。纯化的蛋白酶分子量约为 27 kDa,在 75°C 和 pH 值为 8.0 时具有最佳活性。值得注意的是,该酶即使在没有任何金属离子的情况下也能保持活性,而在 Na+、K+、Mg2+ 和 Cu2+ 金属离子存在的情况下则完全活跃。经测定,其动力学参数的 KM 值为 3.13 mg/ml,Vmax 值为 3.28 × 106 U/mg 。此外,该蛋白酶在处理温度高达 65°C 的环境中持续 60 分钟,以及在 5.0 至 10.0 的广泛 pH 值范围内都表现出了显著的稳定性。这种蛋白酶还能抵御各种恶劣条件,包括有机溶剂、表面活性剂、漂白剂和蛋白水解酶。此外,这种酶在使用商业洗涤剂处理后仍能保持活性,在 4 小时内以 2.50 毫克/毫升的浓度完全溶解血栓。值得注意的是,这种蛋白酶的活性和蛋白质浓度在 4°C 条件下可稳定 70 天。这些发现强调了来自链霉菌 CNXK100 的恒温蛋白酶的潜在工业应用价值。
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引用次数: 0
Isolation and Electrochemical Analysis of a Facultative Anaerobic Electrogenic Strain Klebsiella sp. SQ-1. 兼性厌氧电生菌株克雷伯氏菌 SQ-1 的分离与电化学分析
Pub Date : 2024-04-27 eCollection Date: 2024-06-01 DOI: 10.33073/pjm-2024-013
Lei Zhou, Tuoxian Tang, Dandan Deng, Yayue Wang, Dongli Pei

Electricigens decompose organic matter and convert stored chemical energy into electrical energy through extracellular electron transfer. They are significant biocatalysts for microbial fuel cells with practical applications in green energy generation, effluent treatment, and bioremediation. A facultative anaerobic electrogenic strain SQ-1 is isolated from sludge in a biotechnology factory. The strain SQ-1 is a close relative of Klebsiella variicola. Multilayered biofilms form on the surface of a carbon electrode after the isolated bacteria are inoculated into a microbial fuel cell device. This strain produces high current densities of 625 μA cm-2 by using acetate as the carbon source in a three-electrode configuration. The electricity generation performance is also analyzed in a dual-chamber microbial fuel cell. It reaches a maximum power density of 560 mW m-2 when the corresponding output voltage is 0.59 V. The facultative strain SQ-1 utilizes hydrous ferric oxide as an electron acceptor to perform extracellular electricigenic respiration in anaerobic conditions. Since facultative strains possess better properties than anaerobic strains, Klebsiella sp. SQ-1 may be a promising exoelectrogenic strain for applications in microbial electrochemistry.

电原能分解有机物,并通过细胞外电子传递将储存的化学能转化为电能。它们是微生物燃料电池的重要生物催化剂,可实际应用于绿色能源生产、污水处理和生物修复。从一家生物技术工厂的污泥中分离出了一株兼性厌氧电源菌株 SQ-1。菌株 SQ-1 是变异克雷伯氏菌的近亲。将分离出的细菌接种到微生物燃料电池装置后,碳电极表面形成了多层生物膜。该菌株在三电极配置中使用醋酸盐作为碳源,可产生 625 μA cm-2 的高电流密度。此外,还分析了双室微生物燃料电池的发电性能。兼性菌株 SQ-1 利用含水氧化铁作为电子受体,在厌氧条件下进行胞外电呼吸。由于兼性菌株比厌氧菌株具有更好的特性,因此克雷伯氏菌 SQ-1 可能是微生物电化学应用中一种很有前途的外电源菌株。
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引用次数: 0
Sugar Utilization-Associated Food-Grade Selection Markers in Lactic Acid Bacteria and Yeast. 乳酸菌和酵母中与糖利用相关的食品级选择标记。
Pub Date : 2024-03-04 eCollection Date: 2024-03-01 DOI: 10.33073/pjm-2024-011
Zhiwen Liang, Ke Zheng, Guifeng Xie, Xiongsheng Luo, Huangjin Li

This comprehensive review explores the development of food-grade selection markers in lactic acid bacteria and yeast; some of their strains are precisely defined as safe microorganisms and are crucial in the food industry. Lactic acid bacteria, known for their ability to ferment carbohydrates into lactic acid, provide essential nutrients and contribute to immune responses. With its strong fermentation capabilities and rich nutritional profile, yeast finds use in various food products. Genetic engineering in these microorganisms has grown rapidly, enabling the expression of enzymes and secondary products for food production. However, the focus is on ensuring safety, necessitating food-grade selection markers. Traditional antibiotic and heavy metal resistance selection markers pose environmental and health risks, prompting the search for safer alternatives. Complementary selection markers, such as sugar utilization markers, offer a promising solution. These markers use carbohydrates as carbon sources for growth and are associated with the natural metabolism of lactic acid bacteria and yeast. This review discusses the use of specific sugars, such as lactose, melibiose, sucrose, D-xylose, glucosamine, and N-acetylglucosamine, as selection markers, highlighting their advantages and limitations. In summary, this review underscores the importance of food-grade selection markers in genetic engineering and offers insights into their applications, benefits, and challenges, providing valuable information for researchers in the field of food microbiology and biotechnology.

本综述探讨了乳酸菌和酵母中食品级选择标记的开发;其中一些菌株被精确定义为安全微生物,在食品工业中至关重要。乳酸菌以其将碳水化合物发酵成乳酸的能力而闻名,可提供人体必需的营养物质并促进免疫反应。酵母具有强大的发酵能力和丰富的营养成分,可用于各种食品。这些微生物的基因工程发展迅速,能够表达用于食品生产的酶和次生产品。然而,重点在于确保安全,这就需要食品级的选择标记。传统的抗生素和重金属抗性选择标记会对环境和健康造成危害,因此需要寻找更安全的替代品。补充性选择标记,如糖利用标记,提供了一个很有前景的解决方案。这些标记利用碳水化合物作为生长的碳源,与乳酸菌和酵母菌的自然新陈代谢有关。本综述讨论了特定糖类(如乳糖、瓜寡糖、蔗糖、D-木糖、葡糖胺和 N-乙酰葡糖胺)作为选择标记的使用情况,强调了它们的优势和局限性。总之,这篇综述强调了食品级选择标记在基因工程中的重要性,并深入探讨了它们的应用、益处和挑战,为食品微生物学和生物技术领域的研究人员提供了宝贵的信息。
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引用次数: 0
Association between Clinical Characteristics and Microbiota in Bronchiectasis Patients Based on Metagenomic Next-Generation Sequencing Technology. 基于元基因组新一代测序技术的支气管扩张症患者临床特征与微生物群之间的关系
Pub Date : 2024-03-04 eCollection Date: 2024-03-01 DOI: 10.33073/pjm-2024-007
Dongfeng Shen, Xiaodong Lv, Hui Zhang, Chunyuan Fei, Jing Feng, Jiaqi Zhou, Linfeng Cao, Ying Ying, Na Li, Xiaolong Ma

This study aimed to investigate the disparities between metagenomic next-generation sequencing (mNGS) and conventional culture results in patients with bronchiectasis. Additionally, we sought to investigate the correlation between the clinical characteristics of patients and their microbiome profiles. The overarching goal was to enhance the effective management and treatment of bronchiectasis patients, providing a theoretical foundation for healthcare professionals. A retrospective survey was conducted on 67 bronchiectasis patients admitted to The First Hospital of Jiaxing from October 2019 to March 2023. Clinical baseline information, inflammatory indicators, and pathogen detection reports, including mNGS, conventional blood culture, bronchoalveolar lavage fluid (BALF) culture, and sputum culture results, were collected. By comparing the results of mNGS and conventional culture, the differences in pathogen detection rate and pathogen types were explored, and the diagnostic performance of mNGS compared to conventional culture was evaluated. Based on the various pathogens detected by mNGS, the association between clinical characteristics of bronchiectasis patients and mNGS microbiota results was analyzed. The number and types of pathogens detected by mNGS were significantly larger than those detected by conventional culture. The diagnostic efficacy of mNGS was significantly superior to conventional culture for all types of pathogens, particularly in viral detection (p < 0.01). Regarding pathogen detection rate, the bacteria with the highest detection rate were Pseudomonas aeruginosa (17/58) and Haemophilus influenzae (11/58); the fungus with the highest detection rate was Aspergillus fumigatus (10/21), and the virus with the highest detection rate was human herpes virus 4 (4/11). Differences were observed between the positive and negative groups for P. aeruginosa in terms of common scoring systems for bronchiectasis and whether the main symptom of bronchiectasis manifested as thick sputum (p < 0.05). Significant distinctions were also noted between the positive and negative groups for A. fumigatus regarding Reiff score, neutrophil percentage, bronchiectasis etiology, and alterations in treatment plans following mNGS results reporting (p < 0.05). Notably, 70% of patients with positive A. fumigatus infection opted to change their treatment plans. The correlation study between clinical characteristics of bronchiectasis patients and mNGS microbiological results revealed that bacteria, such as P. aeruginosa, and fungi, such as A. fumigatus, were associated with specific clinical features of patients. This underscored the significance of mNGS in guiding personalized treatment approaches. mNGS could identify multiple pathogens in different types of bronchiectasis samples and was a rapid and effective diagnostic tool for pathogen identification. Its use was recommended for

本研究旨在调查支气管扩张症患者的元基因组下一代测序(mNGS)结果与传统培养结果之间的差异。此外,我们还试图研究患者的临床特征与其微生物组特征之间的相关性。我们的总体目标是加强支气管扩张症患者的有效管理和治疗,为医护人员提供理论基础。研究人员对嘉兴市第一医院2019年10月至2023年3月收治的67名支气管扩张症患者进行了回顾性调查。收集临床基线资料、炎症指标和病原体检测报告,包括mNGS、常规血培养、支气管肺泡灌洗液(BALF)培养和痰培养结果。通过比较 mNGS 和传统培养的结果,探讨了病原体检出率和病原体类型的差异,并评估了 mNGS 与传统培养相比的诊断性能。根据 mNGS 检测出的各种病原体,分析了支气管扩张患者的临床特征与 mNGS 微生物群结果之间的关联。mNGS 检测到的病原体数量和种类明显多于传统培养法检测到的病原体。对于所有类型的病原体,mNGS 的诊断效果都明显优于传统培养,尤其是在病毒检测方面(p < 0.01)。在病原体检出率方面,检出率最高的细菌是铜绿假单胞菌(17/58)和流感嗜血杆菌(11/58);检出率最高的真菌是烟曲霉(10/21);检出率最高的病毒是人类疱疹病毒 4(4/11)。铜绿假单胞菌阳性组和阴性组在支气管扩张常见评分系统和支气管扩张主要症状是否表现为痰液粘稠方面存在差异(P < 0.05)。烟曲霉菌阳性组和阴性组在 Reiff 评分、中性粒细胞百分比、支气管扩张症病因以及 mNGS 结果报告后治疗方案的改变方面也存在显著差异(P < 0.05)。值得注意的是,70% 的烟曲霉菌感染阳性患者选择改变治疗方案。支气管扩张症患者临床特征与 mNGS 微生物结果之间的相关性研究显示,铜绿假单胞菌等细菌和烟曲霉等真菌与患者的特定临床特征相关。mNGS 能在不同类型的支气管扩张样本中鉴定出多种病原体,是一种快速有效的病原体鉴定诊断工具。建议将其用于诊断支气管扩张症患者的感染原因。
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引用次数: 0
Establishment of RPA-Cas12a-Based Fluorescence Assay for Rapid Detection of Feline Parvovirus. 建立基于 RPA-Cas12a 的荧光检测方法,用于快速检测猫细小病毒。
Pub Date : 2024-03-04 eCollection Date: 2024-03-01 DOI: 10.33073/pjm-2024-005
Ting Wang, Hao Zeng, Qiming Liu, Weidong Qian, Yongdong Li, Jian Liu, Rong Xu

Feline parvovirus (FPV) is highly infectious for cats and other Felidae and often causes severe damage to young kittens. In this study, we incorporated recombinase polymerase amplification (RPA) and Cas12a-mediated detection and developed an RPA-Cas12a-based real-time or end-point fluorescence detection method to identify the NS1 gene of FPV. The total time of RPA-Cas12a-based fluorescence assay is approximately 25 min. The assay presented a limit of detection (LOD) of 1 copies/μl (25 copies/per reaction), with no cross-reactivity with several feline pathogens. The clinical performance of the assay was examined using total genomic DNA purified from 60 clinical specimens and then compared to results obtained with qPCR detection of FPV with 93.3% positive predictive agreement and 100% negative predictive agreement. Together, the rapid reaction, cost-effectiveness, and high sensitivity make the RPA-Cas12a-based fluorescence assay a fascinating diagnostic tool that will help minimize infection spread through instant detection of FPV.

猫细小病毒(FPV)对猫和其他猫科动物具有高度传染性,经常对幼猫造成严重伤害。本研究结合重组酶聚合酶扩增(RPA)和Cas12a介导的检测方法,开发了一种基于RPA-Cas12a的实时或终点荧光检测方法来鉴定FPV的NS1基因。基于 RPA-Cas12a 的荧光检测总时间约为 25 分钟。该检测方法的检测限(LOD)为1拷贝/μl(25拷贝/次反应),与多种猫科病原体无交叉反应。使用从 60 份临床标本中纯化的总基因组 DNA 检验了该检测方法的临床性能,并将其与 qPCR 检测 FPV 的结果进行了比较,两者的阳性预测一致率为 93.3%,阴性预测一致率为 100%。基于 RPA-Cas12a 的荧光检测具有反应迅速、成本效益高和灵敏度高等特点,是一种极具吸引力的诊断工具,有助于通过即时检测 FPV 最大限度地减少感染传播。
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引用次数: 0
A Mendelian Randomization Study: Roles of Gut Microbiota in Sepsis - Who is the Angle? 孟德尔随机研究:肠道微生物群在败血症中的作用--谁是角度?
Pub Date : 2024-03-04 eCollection Date: 2024-03-01 DOI: 10.33073/pjm-2024-006
Yeping Bian, Jian Xu, Xiaojing Deng, Suming Zhou

Gut microbiota (GM) is a crucial underlying player during sepsis pathogenesis. However, the causal relationship is unclear and remains to be determined. A two-sample Mendelian randomization study was implemented. The statistical data about sepsis together with GM summarized from genome-wide association studies were evaluated. Instrumental variables were defined as single-nucleotide polymorphisms with prominent correlations with exposure. The inverse-variance-weighted test was employed as a major approach of Mendelian randomization analysis to estimate of causal relationships. The inverse-variance-weighted analysis results demonstrated that at different taxa levels, Actinobacteria and Bifidobacteriaceae influence sepsis. Actinobacteria had negative relationships to sepsis risk at the phylum (β = -0.34, SE = 0.10, p = 0.0008) and class (β = -0.23, SE = 0.07, p = 0.0011) levels in outcome coded ieu-b-69. Actinobacteria at the phylum level (β = -0.22, SE = 0.10, p = 0.027) was also negatively associated with sepsis in outcome coded ieu-b-4980. Bifidobacteriaceae at the order (β = -0.20, SE = 0.06, p = 0.0021), family (β = -0.20, SE = 0.06, p = 0.0021), and genus (β = -0.20, SE = 0.06, p = 0.0007) levels were all negatively correlated with the risk of sepsis in outcome coded ieu-b-69. The results of the Wald ratio model showed that Tyzzerella genus (OR (95%CI) = 0.6902[0.4907,0.9708], p = 0.0331) and Gastranaerophilales order (OR (95%CI) = 0.5907[0.3516,0.9926], p = 0.0468) were negatively connected with sepsis. This study implied at different taxa levels Actinobacteria and Bifidobacteriaceae, Tyzzerella genus, and Gastranaerophilales order have a causal relationship with sepsis, indicating that they are protective factors for the incidence of sepsis.

肠道微生物群(GM)是脓毒症发病机制中一个至关重要的潜在因素。然而,其因果关系尚不明确,仍有待确定。我们进行了一项双样本孟德尔随机研究。研究人员评估了从全基因组关联研究中总结的有关脓毒症和 GM 的统计数据。工具变量被定义为与暴露有显著相关性的单核苷酸多态性。反方差加权检验作为孟德尔随机分析的一种主要方法,被用来估计因果关系。逆方差加权分析结果表明,在不同类群水平上,放线菌和双歧杆菌科对败血症有影响。在编码为 ieu-b-69 的结果中,放线菌在门级(β = -0.34,SE = 0.10,p = 0.0008)和类级(β = -0.23,SE = 0.07,p = 0.0011)与败血症风险呈负相关。在编码为 ieu-b-4980 的结果中,门级放线菌(β = -0.22,SE = 0.10,p = 0.027)也与败血症呈负相关。双歧杆菌科的目级(β = -0.20,SE = 0.06,p = 0.0021)、科级(β = -0.20,SE = 0.06,p = 0.0021)和属级(β = -0.20,SE = 0.06,p = 0.0007)均与结果编码为 ieu-b-69 的败血症风险呈负相关。Wald ratio 模型的结果显示,Tyzzerella 属(OR (95%CI) = 0.6902[0.4907,0.9708], p = 0.0331)和 Gastranaerophilales 目(OR (95%CI) = 0.5907[0.3516,0.9926], p = 0.0468)与败血症呈负相关。这项研究表明,在不同类群水平上,放线菌和双歧杆菌科、Tyzzerella属和嗜胃肠菌目与败血症有因果关系,表明它们是败血症发病率的保护因素。
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引用次数: 0
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Polish journal of microbiology
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