Pub Date : 2024-11-05DOI: 10.1016/j.actatropica.2024.107451
Hasan Y. Alniss , Sreedevi Sajeev , Ruqaiyyah Siddiqui , Meshal Daalah , Bader S. Alawfi , Hadeel M. Al-Jubeh , Anil Ravi , Naveed A. Khan
DNA minor groove binders exhibit a high degree of sequence specificity and have a variety of biological actions including antiviral, anticancer, antibacterial, and anti-protozoal properties. Since it is the location of non-covalent interactions, the minor groove of double helical B-DNA is gaining significant interest as therapeutic targets. For the purpose of this investigation, the synthesis of five novel DNA minor groove binding agents was accomplished and antiparasitic efficacies were determined against Acanthamoeba castellanii of the T4 genotype in vitro. Using amoebicidal assays, the results revealed that all inhibitors tested showed significant killing of amoebae (P < 0.05). Pre-treatment of amoebae with DNA minor groove binders inhibited parasite-mediated human cell death by measuring lactate dehydrogenase release using cytopathogenicity assays. Cytotoxicity assays revealed minimal effects on human cells. As phenotypic switching leads to infection recurrence, assays revealed that inhibitors blocked amoebae phenotypic transformation. These are promising findings and suggest that DNA minor groove binders may hold promise for further research in the effective eradication of pathogenic A. castellanii.
DNA 小沟结合剂具有高度的序列特异性，并具有多种生物作用，包括抗病毒、抗癌、抗菌和抗原虫特性。由于它是非共价相互作用的位置，双螺旋 B-DNA 的小沟作为治疗靶点正受到越来越多的关注。为了进行这项研究，我们合成了五种新型 DNA 小沟结合剂，并在体外测定了它们对 T4 基因型卡斯特阿米巴的抗寄生虫效果。通过变形虫杀灭试验，结果表明所有测试的抑制剂都能显著杀灭变形虫（P
{"title":"Targeting pathogenic Acanthamoeba castellanii using DNA minor groove binding agents","authors":"Hasan Y. Alniss , Sreedevi Sajeev , Ruqaiyyah Siddiqui , Meshal Daalah , Bader S. Alawfi , Hadeel M. Al-Jubeh , Anil Ravi , Naveed A. Khan","doi":"10.1016/j.actatropica.2024.107451","DOIUrl":"10.1016/j.actatropica.2024.107451","url":null,"abstract":"<div><div>DNA minor groove binders exhibit a high degree of sequence specificity and have a variety of biological actions including antiviral, anticancer, antibacterial, and anti-protozoal properties. Since it is the location of non-covalent interactions, the minor groove of double helical B-DNA is gaining significant interest as therapeutic targets. For the purpose of this investigation, the synthesis of five novel DNA minor groove binding agents was accomplished and antiparasitic efficacies were determined against <em>Acanthamoeba castellanii</em> of the T4 genotype <em>in vitro</em>. Using amoebicidal assays, the results revealed that all inhibitors tested showed significant killing of amoebae (<em>P</em> < 0.05). Pre-treatment of amoebae with DNA minor groove binders inhibited parasite-mediated human cell death by measuring lactate dehydrogenase release using cytopathogenicity assays. Cytotoxicity assays revealed minimal effects on human cells. As phenotypic switching leads to infection recurrence, assays revealed that inhibitors blocked amoebae phenotypic transformation. These are promising findings and suggest that DNA minor groove binders may hold promise for further research in the effective eradication of pathogenic <em>A. castellanii</em>.</div></div>","PeriodicalId":7240,"journal":{"name":"Acta tropica","volume":"260 ","pages":"Article 107451"},"PeriodicalIF":2.1,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142602562","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-05DOI: 10.1016/j.actatropica.2024.107454
Rafael A.N. Ramos , Hassan Hakimi , Sidouin K. Metinou , Wachellet Danzabe , Macon Overcast , Jeremiah Cox , Rebecca Garabed , Philip Tchindebet Ouakou , Richard Ngandolo Bongo Nare , Fernando Torres-Velez , Lucienne Tritten , Meriam N. Saleh , Guilherme G. Verocai
Cutaneous myiasis caused by various Calliphoridae dipteran species is prevalent worldwide and is of particular veterinary and public health concern. Recently, in a scientific exploration of the Guinea Worm Eradication Program to Chad, Africa, we observed that dogs with mutilated ears, based on local awareness, were caused by cutaneous myiasis. In this study, we analyzed epidemiological, morphological, and molecular data on cutaneous myiasis in dogs from Chad. From September to October 2022, dogs (n = 1,562) from 56 villages situated along the Chari River were physically inspected for cutaneous myiasis. All larvae were collected and identified morphologically and by molecular analysis of the partial cytochrome oxidase c subunit I (COI) mitochondrial gene. The prevalence of myiasis infestation along with 95 % confidence intervals (95 % CI) was determined using the modified Wilson method. Myiasis was detected in dogs from 21 villages (37.5 %; 95 % CI 26 – 50 %), predominating in the southernmost region. Of 1,562 dogs, 66 (4.22 %; 95 % CI 3.34 – 5.34 %) were infested by calliphorid larvae, with a mean infestation of 2.28 larvae per animal (range = 1 to 24). Specimens were morphologically identified as Cordylobia anthropophaga (n = 94), Chrysomya bezziana (n = 54), and Chrysomya sp. (n = 3), which were detected in 57, eight and one dog, respectively. No co-infestations were observed. The molecular analyses confirmed the morphological identification and revealed the presence of 17 haplotypes for C. anthropophaga, 2 for C. bezziana, and one for Chrysomya sp. Our study emphasizes the veterinary importance of myiasis in dogs in Africa and proposes measures to assure their health and well-being.
{"title":"Cutaneous myiasis by Calliphoridae dipterans in dogs from Chad","authors":"Rafael A.N. Ramos , Hassan Hakimi , Sidouin K. Metinou , Wachellet Danzabe , Macon Overcast , Jeremiah Cox , Rebecca Garabed , Philip Tchindebet Ouakou , Richard Ngandolo Bongo Nare , Fernando Torres-Velez , Lucienne Tritten , Meriam N. Saleh , Guilherme G. Verocai","doi":"10.1016/j.actatropica.2024.107454","DOIUrl":"10.1016/j.actatropica.2024.107454","url":null,"abstract":"<div><div>Cutaneous myiasis caused by various Calliphoridae dipteran species is prevalent worldwide and is of particular veterinary and public health concern. Recently, in a scientific exploration of the Guinea Worm Eradication Program to Chad, Africa, we observed that dogs with mutilated ears, based on local awareness, were caused by cutaneous myiasis. In this study, we analyzed epidemiological, morphological, and molecular data on cutaneous myiasis in dogs from Chad. From September to October 2022, dogs (<em>n</em> = 1,562) from 56 villages situated along the Chari River were physically inspected for cutaneous myiasis. All larvae were collected and identified morphologically and by molecular analysis of the partial cytochrome oxidase c subunit I (COI) mitochondrial gene. The prevalence of myiasis infestation along with 95 % confidence intervals (95 % CI) was determined using the modified Wilson method. Myiasis was detected in dogs from 21 villages (37.5 %; 95 % CI 26 – 50 %), predominating in the southernmost region. Of 1,562 dogs, 66 (4.22 %; 95 % CI 3.34 – 5.34 %) were infested by calliphorid larvae, with a mean infestation of 2.28 larvae per animal (range = 1 to 24). Specimens were morphologically identified as <em>Cordylobia anthropophaga</em> (<em>n</em> = 94), <em>Chrysomya bezziana</em> (<em>n</em> = 54), and <em>Chrysomya</em> sp. (<em>n</em> = 3), which were detected in 57, eight and one dog, respectively. No co-infestations were observed. The molecular analyses confirmed the morphological identification and revealed the presence of 17 haplotypes for <em>C. anthropophaga</em>, 2 for <em>C. bezziana</em>, and one for <em>Chrysomya</em> sp. Our study emphasizes the veterinary importance of myiasis in dogs in Africa and proposes measures to assure their health and well-being.</div></div>","PeriodicalId":7240,"journal":{"name":"Acta tropica","volume":"260 ","pages":"Article 107454"},"PeriodicalIF":2.1,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142589644","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-05DOI: 10.1016/j.actatropica.2024.107455
Reynoso Mercedes María Noel , Alzogaray Raúl Adolfo
Eugenol is a botanical monoterpene found in the essential oils of several aromatic plants. It has shown to have insecticidal activity, modify insect behavior, and its site of action is most probably in the octopaminergic system. The aim of the present study was to explore whether tyramine receptors are involved in the hyperactivity produced by eugenol in Triatoma infestans, one of the main vectors of Chagas disease. Topical application of tyramine and eugenol increased locomotor activity in third instar nymphs of T. infestans in a dose-dependent way. Yohimbine hydrochloride, a tyramine antagonist, did not modify nymph activity. However, nymphs pretreated with yohimbine and then with tyramine or eugenol showed the same locomotor activity as the controls. Therefore, yohimbine hydrochloride inhibited the hyperactivating effect of both tyramine and eugenol. These results suggest that tyramine receptors are involved in the effect of eugenol on the locomotor activity of T. infestans.
{"title":"An antagonist of the neurotransmitter tyramine inhibits the hyperactivating effect of eugenol in the blood-sucking bug, Triatoma infestans","authors":"Reynoso Mercedes María Noel , Alzogaray Raúl Adolfo","doi":"10.1016/j.actatropica.2024.107455","DOIUrl":"10.1016/j.actatropica.2024.107455","url":null,"abstract":"<div><div>Eugenol is a botanical monoterpene found in the essential oils of several aromatic plants. It has shown to have insecticidal activity, modify insect behavior, and its site of action is most probably in the octopaminergic system. The aim of the present study was to explore whether tyramine receptors are involved in the hyperactivity produced by eugenol in <em>Triatoma infestans</em>, one of the main vectors of Chagas disease. Topical application of tyramine and eugenol increased locomotor activity in third instar nymphs of <em>T. infestans</em> in a dose-dependent way. Yohimbine hydrochloride, a tyramine antagonist, did not modify nymph activity. However, nymphs pretreated with yohimbine and then with tyramine or eugenol showed the same locomotor activity as the controls. Therefore, yohimbine hydrochloride inhibited the hyperactivating effect of both tyramine and eugenol. These results suggest that tyramine receptors are involved in the effect of eugenol on the locomotor activity of <em>T. infestans</em>.</div></div>","PeriodicalId":7240,"journal":{"name":"Acta tropica","volume":"260 ","pages":"Article 107455"},"PeriodicalIF":2.1,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142602564","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-02DOI: 10.1016/j.actatropica.2024.107453
Carol Sanchez-Chicana , Lisseth M. Leiva , Juan Jimenez-Chunga , Walter Silva , Javier Jara , Teresa Lopez-Urbina , Armando E. Gonzalez , Miguel Rojas , Luis A. Gomez-Puerta
Coronaviruses are common around the world and infect a wide variety of animals, including domestic and wild ones. They are characterized by causing respiratory, enteric, hepatic, and neurological diseases of varying severity, from asymptomatic to severe. Wild animals play a crucial role in this group of viruses since they can act as hosts or reservoirs for pathogenic species of humans and domestic animals. The purpose of this study was to molecularly identify coronaviruses present in wild mammals seized and rescued by the National Forestry and Wildlife Service (SERFOR) of Peru. We molecularly analyzed tracheal and rectal swabs from 90 wild mammals seized and/or rescued by SERFOR, partially amplifying the coronavirus RdRp gene. Ten of the 90 animals studied (11.1%) were positive only for Alphacoronavirus. These were non-human primates (Aotus sp., Sapajus apella, and Saimiri sciureus), the crab-eating raccoon (Procyon cancrivorus), and the South American sea lion (Otaria flavescens). The partial sequence analysis of the RdRp gene revealed that nine sequences belonged to the Pedacovirus subgenus and shared 99.1% nucleotide identity with the porcine epidemic diarrhea virus (PEDV), and only one sequence belonged to the Tegacovirus subgenus and shared 95.6% identity with the Feline coronavirus (FCoV). The results show that various wild mammal species from Peru can act as hosts for coronaviruses capable of infecting domestic species. Due to this, it is necessary to implement measures that help us identify the genera and species of coronaviruses in these species to prevent and contain future epidemics or pandemics resulting from the high rate of recombination and mutation of this virus.
{"title":"Surveillance of coronavirus in wild mammals seized and rescued by the National Forest and Wildlife Service of Peru","authors":"Carol Sanchez-Chicana , Lisseth M. Leiva , Juan Jimenez-Chunga , Walter Silva , Javier Jara , Teresa Lopez-Urbina , Armando E. Gonzalez , Miguel Rojas , Luis A. Gomez-Puerta","doi":"10.1016/j.actatropica.2024.107453","DOIUrl":"10.1016/j.actatropica.2024.107453","url":null,"abstract":"<div><div>Coronaviruses are common around the world and infect a wide variety of animals, including domestic and wild ones. They are characterized by causing respiratory, enteric, hepatic, and neurological diseases of varying severity, from asymptomatic to severe. Wild animals play a crucial role in this group of viruses since they can act as hosts or reservoirs for pathogenic species of humans and domestic animals. The purpose of this study was to molecularly identify coronaviruses present in wild mammals seized and rescued by the National Forestry and Wildlife Service (SERFOR) of Peru. We molecularly analyzed tracheal and rectal swabs from 90 wild mammals seized and/or rescued by SERFOR, partially amplifying the coronavirus RdRp gene. Ten of the 90 animals studied (11.1%) were positive only for <em>Alphacoronavirus</em>. These were non-human primates (<em>Aotus</em> sp., <em>Sapajus apella</em>, and <em>Saimiri sciureus</em>), the crab-eating raccoon (<em>Procyon cancrivorus</em>), and the South American sea lion (<em>Otaria flavescens</em>). The partial sequence analysis of the RdRp gene revealed that nine sequences belonged to the <em>Pedacovirus</em> subgenus and shared 99.1% nucleotide identity with the porcine epidemic diarrhea virus (PEDV), and only one sequence belonged to the <em>Tegacovirus</em> subgenus and shared 95.6% identity with the Feline coronavirus (FCoV). The results show that various wild mammal species from Peru can act as hosts for coronaviruses capable of infecting domestic species. Due to this, it is necessary to implement measures that help us identify the genera and species of coronaviruses in these species to prevent and contain future epidemics or pandemics resulting from the high rate of recombination and mutation of this virus.</div></div>","PeriodicalId":7240,"journal":{"name":"Acta tropica","volume":"260 ","pages":"Article 107453"},"PeriodicalIF":2.1,"publicationDate":"2024-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142566284","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Theileria parasites were investigated from cattle ticks (Rhipicephalus microplus (Canestrini, 1888)) collected in 12 provinces in upper-northeastern Thailand based on the sequences of 18S rRNA and MPSP gene. Polymerase chain reactions (PCRs) and sequencing specific regions for the 18S rRNA gene revealed two species of Theileria pathogens; T. orientalis (n = 42) and T. sinensis (n = 31) with 94.50−100 % identity. In the provinces of upper-northeastern Thailand, the nucleotide diversity of Theileria's 18S rRNA for T. sinensis and T. orientalis were 0 % and 1.3 %. respectively. The MPSP gene was used to categorize the T. orientalis genotypes. The sequences were compared with those available in the public database (GenBank) for species identification. Phylogenetic trees of Theileria were constructed from the MPSP gene sequences of our amplicons and those available in GenBank using maximum-likelihood and neighbor-joining analyses. The results revealed three identified genotypes: type 3, 5, and 7. Although the main carriers of T. orientalis are ticks in the genus Haemaphysalis, T. orientalis was the most frequently found in R. microplus in upper-northeastern Thailand. Theileria was frequent in Nong Khai, Mukdahan, and Loei, three Thai provinces that bordered the Lao PDR close to the Mekong River. Epidemiological surveys and control strategies in this region should be considered.
根据 18S rRNA 和 MPSP 基因的序列，研究了从泰国东北部上游 12 个府采集的牛虱（Rhipicephalus microplus (Canestrini, 1888)）中的寄生虫。聚合酶链式反应（PCR）和 18S rRNA 基因特定区域的测序结果显示，病原菌有两个物种：T. orientalis（n=42）和 T. sinensis（n=31），其同一性为 94.50%-100%。在泰国上东北部各府，T. sinensis 和 T. orientalis 的丝核菌 18S rRNA 核苷酸多样性分别为 0% 和 1.3%。利用 MPSP 基因对东方蓟马的基因型进行了分类。将序列与公共数据库（GenBank）中的序列进行比较，以确定物种。使用最大似然法和邻接分析法，根据我们的扩增子的 MPSP 基因序列和 GenBank 中的序列构建了东方蓟马的系统发生树。结果发现了三种已确定的基因型：3 型、5 型和 7 型。虽然 T. orientalis 的主要携带者是 Haemaphysalis 属的蜱虫，但在泰国上东北部的 R. microplus 中最常发现 T. orientalis。在靠近湄公河、与老挝人民民主共和国接壤的泰国廊开府、木爹罕府和乐毅府，东方蜱属螨虫十分常见。应考虑在这一地区开展流行病学调查并制定控制策略。
{"title":"Detection of Theileria in cattle ticks (Rhipicephalus microplus) (Canestrini, 1888) in upper-northeastern Thailand","authors":"Kanchana Thinnabut , Rutchanee Rodpai , Oranuch Sanpool , Wanchai Maleewong , Ubon Tangkawanit","doi":"10.1016/j.actatropica.2024.107452","DOIUrl":"10.1016/j.actatropica.2024.107452","url":null,"abstract":"<div><div><em>Theileria</em> parasites were investigated from cattle ticks (<em>Rhipicephalus microplus</em> (Canestrini, 1888)) collected in 12 provinces in upper-northeastern Thailand based on the sequences of 18S rRNA and MPSP gene. Polymerase chain reactions (PCRs) and sequencing specific regions for the 18S rRNA gene revealed two species of <em>Theileria</em> pathogens; <em>T. orientalis</em> (n = 42) and <em>T. sinensis</em> (n = 31) with 94.50−100 % identity. In the provinces of upper-northeastern Thailand, the nucleotide diversity of Theileria's 18S rRNA for <em>T. sinensis</em> and <em>T. orientalis</em> were 0 % and 1.3 %. respectively. The MPSP gene was used to categorize the <em>T. orientalis</em> genotypes. The sequences were compared with those available in the public database (GenBank) for species identification. Phylogenetic trees of <em>Theileria</em> were constructed from the MPSP gene sequences of our amplicons and those available in GenBank using maximum-likelihood and neighbor-joining analyses. The results revealed three identified genotypes: type 3, 5, and 7. Although the main carriers of <em>T. orientalis</em> are ticks in the genus <em>Haemaphysalis, T. orientalis</em> was the most frequently found in <em>R. microplus</em> in upper-northeastern Thailand. <em>Theileria</em> was frequent in Nong Khai, Mukdahan, and Loei, three Thai provinces that bordered the Lao PDR close to the Mekong River. Epidemiological surveys and control strategies in this region should be considered.</div></div>","PeriodicalId":7240,"journal":{"name":"Acta tropica","volume":"260 ","pages":"Article 107452"},"PeriodicalIF":2.1,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142567273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-01DOI: 10.1016/j.actatropica.2024.107450
Thaysa Carolina Gonçalves Silva , Paula Carolina Valença Silva , Elker Lene Santos de Lima , Maria Tereza Cartaxo Muniz , Edmundo Pessoa Lopes , Ana Lúcia Coutinho Domingues
In schistosomiasis mansoni (SM), periportal fibrosis (PPF) arises due to an inflammatory response exacerbated by parasite eggs in the intrahepatic portal space, culminating in the deposition of collagen and extracellular matrix proteins. This fibrosis results from a remodeling process of the extracellular matrix, in which metalloproteinases play a significant role. The study evaluated the association between MMP-3 polymorphism (-1171 5A>6A) (rs 3025058) and sociodemographic factors with PPF in individuals with SM. This is an analytical cross-sectional study involving 242 individuals infected with S. mansoni, of these 122 were diagnosed with hepatosplenic form (HS) and 119 hepatointestinal form (HI), all from the state of Pernambuco, Brazil. Polymerase chain reaction with restriction enzyme digestion (Psyl) was used to determine the MMP-3 polymorphism (-1171 5A>6A). There was a significant association between the male gender and the HS form (OR = 1.7623 95% CI [1.0481–2.9631]; p-value = 0.0439) as well as individuals aged over 41, also had a greater chance of developing this clinical form of the disease (OR = 2.8299; 95% CI [1.5211–5.2650]; p-value = 0.0014), with greater emphasis on individuals over 61 years old (OR= 8.5541; 95% CI [3.6895–19.8326], p-value= 0.0000). There was no statistically significant association between the MMP-3 polymorphism (-1171 5A>6A) between the clinical groups (5A6A CI [0.7144–1.9879] p-value 0.5882 5A5A CI [0.0912–2.9231] p-value 0.7331 5A6A / 5A5A CI [0.6904–1.8937] p-value 0.6949). In conclusion, the results showed no association between the MMP-3 polymorphism (-1171 5A>6A) and the development of PPF. In addition, males, and age over 41 were predictive factors for the HS form of the disease in this Brazilian population.
在曼氏血吸虫病（SM）中，肝门静脉周围纤维化（PPF）是由于肝门静脉内寄生虫卵加剧了炎症反应，最终导致胶原蛋白和细胞外基质蛋白沉积。这种纤维化是细胞外基质重塑过程的结果，金属蛋白酶在其中发挥了重要作用。本研究评估了 SM 患者的 MMP-3 多态性（-1171 5A>6A）（rs 3025058）和社会人口因素与 PPF 之间的关联。这是一项横断面分析研究，涉及 242 名曼氏沙门氏菌感染者，其中 122 人被诊断为肝脾型（HS），119 人被诊断为肝肠型（HI），他们都来自巴西伯南布哥州。聚合酶链反应与限制性酶消化（Psyl）被用来确定 MMP-3 的多态性（-1171 5A>6A）。男性性别与 HS 形态之间存在明显关联（OR = 1.7623 95% CI [1.0481-2.9631]；P 值 = 0.0439），41 岁以上的人患这种临床形态疾病的几率也更大（OR = 2.8299；95% CI [1.5211-5.2650]；P 值 = 0.0014），61 岁以上者的发病率更高（OR= 8.5541；95% CI [3.6895-19.8326]，P 值 = 0.0000）。临床组之间的 MMP-3 多态性（-1171 5A>6A）之间没有统计学意义上的显著关联（5A6A CI [0.7144-1.9879] p-value 0.5882 5A5A CI [0.0912-2.9231] p-value 0.7331 5A6A / 5A5A CI [0.6904-1.8937] p-value 0.6949）。总之，研究结果表明，MMP-3 多态性（-1171 5A>6A）与 PPF 的发生没有关联。此外，在这一巴西人群中，男性和 41 岁以上是预测 HE 型疾病的因素。
{"title":"Influence of metalloproteinase-3 (-1171 5A>6A) polymorphism on periportal fibrosis in patients with schistosomiasis mansoni, Pernambuco, Brazil","authors":"Thaysa Carolina Gonçalves Silva , Paula Carolina Valença Silva , Elker Lene Santos de Lima , Maria Tereza Cartaxo Muniz , Edmundo Pessoa Lopes , Ana Lúcia Coutinho Domingues","doi":"10.1016/j.actatropica.2024.107450","DOIUrl":"10.1016/j.actatropica.2024.107450","url":null,"abstract":"<div><div>In schistosomiasis mansoni (SM), periportal fibrosis (PPF) arises due to an inflammatory response exacerbated by parasite eggs in the intrahepatic portal space, culminating in the deposition of collagen and extracellular matrix proteins. This fibrosis results from a remodeling process of the extracellular matrix, in which metalloproteinases play a significant role. The study evaluated the association between <em>MMP-3</em> polymorphism (-1171 <em>5A>6A</em>) (rs 3025058) and sociodemographic factors with PPF in individuals with SM. This is an analytical cross-sectional study involving 242 individuals infected with <em>S. mansoni</em>, of these 122 were diagnosed with hepatosplenic form (HS) and 119 hepatointestinal form (HI), all from the state of Pernambuco, Brazil. Polymerase chain reaction with restriction enzyme digestion (Psyl) was used to determine the <em>MMP-3</em> polymorphism (-1171 5A>6A). There was a significant association between the male gender and the HS form (OR = 1.7623 95% CI [1.0481–2.9631]; p-value = 0.0439) as well as individuals aged over 41, also had a greater chance of developing this clinical form of the disease (OR = 2.8299; 95% CI [1.5211–5.2650]; p-value = 0.0014), with greater emphasis on individuals over 61 years old (OR= 8.5541; 95% CI [3.6895–19.8326], p-value= 0.0000). There was no statistically significant association between the <em>MMP-3</em> polymorphism (-1171 5A>6A) between the clinical groups (5A6A CI [0.7144–1.9879] p-value 0.5882 5A5A CI [0.0912–2.9231] p-value 0.7331 5A6A / 5A5A CI [0.6904–1.8937] p-value 0.6949). In conclusion, the results showed no association between the <em>MMP-3</em> polymorphism (-1171 5A>6A) and the development of PPF. In addition, males, and age over 41 were predictive factors for the HS form of the disease in this Brazilian population.</div></div>","PeriodicalId":7240,"journal":{"name":"Acta tropica","volume":"260 ","pages":"Article 107450"},"PeriodicalIF":2.1,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142567278","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-31DOI: 10.1016/j.actatropica.2024.107446
Bandar Hasan Saleh , Allan Lugaajju , Muyideen Kolapo Tijani , Lena Danielsson , Ulrika Morris , Kristina E M Persson
Malaria caused by Plasmodium falciparum leads to the destruction of red blood cells (RBCs). A better understanding of how naturally immune individuals control infections should be valuable for future vaccine studies. Antibodies against RBCs and RBC surface antigens were measured together with different inflammatory markers in healthy adults living in a malaria endemic area of Uganda and compared to Swedish healthy adults. Antibodies binding to RBCs were clearly elevated in Ugandans compared to Swedish samples, and for RBC surface antigens the Ugandans had higher levels of antibodies against JMH, but not against Cromer or Kell. Twenty-eight percent of the Ugandans were PCR-positive for P. falciparum, and these had higher levels of IgG against parasite extract and more inhibition in functional growth/invasion assays, but levels of antibodies against RBC, RBC surface antigens, results from Direct Antiglobulin Tests (DAT) and indirect antiglobulin tests were similar when compared with PCR-negative individuals. When inflammatory markers (α-1-antitrypsin, haptoglobin, orosomucoid/α-1-acid glycoprotein, CRP, IgG, IgA and IgM) were measured there were in general almost no signs of inflammation except for clearly elevated levels of IgG. Some had low levels of haptoglobin and for orosomucoid more than half of the individuals had clearly reduced levels. There was no correlation between the inflammatory markers and PCR-positivity, antibodies against RBCs or parasites. In conclusion, for healthy adults living in a malaria endemic area, there was a clear presence of antibodies against RBCs in parallel with high levels of IgG and almost no signs of inflammation, even though many individuals were carrying parasites.
{"title":"An immuno-inflammatory profiling of asymptomatic individuals in a malaria endemic area in Uganda","authors":"Bandar Hasan Saleh , Allan Lugaajju , Muyideen Kolapo Tijani , Lena Danielsson , Ulrika Morris , Kristina E M Persson","doi":"10.1016/j.actatropica.2024.107446","DOIUrl":"10.1016/j.actatropica.2024.107446","url":null,"abstract":"<div><div>Malaria caused by <em>Plasmodium falciparum</em> leads to the destruction of red blood cells (RBCs). A better understanding of how naturally immune individuals control infections should be valuable for future vaccine studies. Antibodies against RBCs and RBC surface antigens were measured together with different inflammatory markers in healthy adults living in a malaria endemic area of Uganda and compared to Swedish healthy adults. Antibodies binding to RBCs were clearly elevated in Ugandans compared to Swedish samples, and for RBC surface antigens the Ugandans had higher levels of antibodies against JMH, but not against Cromer or Kell. Twenty-eight percent of the Ugandans were PCR-positive for <em>P. falciparum</em>, and these had higher levels of IgG against parasite extract and more inhibition in functional growth/invasion assays, but levels of antibodies against RBC, RBC surface antigens, results from Direct Antiglobulin Tests (DAT) and indirect antiglobulin tests were similar when compared with PCR-negative individuals. When inflammatory markers (α-1-antitrypsin, haptoglobin, orosomucoid/α-1-acid glycoprotein, CRP, IgG, IgA and IgM) were measured there were in general almost no signs of inflammation except for clearly elevated levels of IgG. Some had low levels of haptoglobin and for orosomucoid more than half of the individuals had clearly reduced levels. There was no correlation between the inflammatory markers and PCR-positivity, antibodies against RBCs or parasites. In conclusion, for healthy adults living in a malaria endemic area, there was a clear presence of antibodies against RBCs in parallel with high levels of IgG and almost no signs of inflammation, even though many individuals were carrying parasites.</div></div>","PeriodicalId":7240,"journal":{"name":"Acta tropica","volume":"260 ","pages":"Article 107446"},"PeriodicalIF":2.1,"publicationDate":"2024-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142563729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-30DOI: 10.1016/j.actatropica.2024.107449
Lais Sampaio de Azevedo , Vanessa Cristina Martins Silva , Raquel Guiducci , Simone Guadagnucci , Fernanda Faria Costa , Monique Beerens Abdul Ghani , Ricardo Duarte Lopes , Antonio Charlys da Costa , Lia Cunha , Marcilio Figueredo Lemos , Adriana Parise , Regina Célia Moreira , Adriana Luchs
Recent increases in zoonotic diseases underscore the integration of companion animals into urban environments, posing complex transmission risks and highlighting the necessity of One Health approaches. Respiratory and enteric viruses have been consistently linked to interspecies transmission between humans and animals. This study aimed to assess the circulation of human noroviruses (NoV), human adenoviruses (HAdV), enteroviruses (EV), parechoviruses (PeV-A), human bocaviruses (HBoV), hepatitis A (HAV) and E viruses (HEV), Influenza A and B viruses (Flu A/B), respiratory syncytial virus (RSV), and SARS-CoV-2 in domestic dogs and cats in Brazil to understand potential zooanthroponosis risks. Between 2012 and 2021, 600 fecal samples from dogs and cats (516 and 84, respectively) were collected at small animal clinics in São Paulo state, Brazil. The specimens underwent in-house qPCR screening for HBoV and HAdV, while EV, PeV-A, NoV, and HEV were tested using in-house RT-qPCR. SARS-CoV-2, Flu A/B, and RSV were investigated with a commercial RT-qPCR kit assay. HAV detection utilized conventional nested (RT)-PCR. Positive samples were sequenced for molecular characterization and phylogenetic analysis. NoV was detected in 0.2 % (1/600) of the animals, while all other investigated viruses tested negative. The NoV-positive sample, collected in 2012 from a pet dog, was identified as genotype GII.4_Sydney[P31]. The Dog/BRA/2012/GII.4_Sydney[P31]/IAL-M21 strain exhibited a close genetic relationship to Brazilian human and environmental NoV GII.4_Sydney[P31] strains, with 98.1–99.2 % nucleotide similarity in ORF1 and 99.2–99.6 % in ORF2 sequences, suggesting interspecies transmission. Pet dogs are frequently exposed to human fecal-borne viruses, highlighting the potential for zooanthroponotic transmission due to their close interaction with humans in shared environments. There is an urgent need to enhance surveillance studies in companion animals to better understand the implications of detecting human NoV strains in pets, as NoV could potentially act as a reverse zoonotic disease in households, animal hospitals, or shelters worldwide.
{"title":"Emerging zooanthroponotic risks: Detection of the human norovirus GII.4 Sydney[P31] strain in a domestic dog in Brazil","authors":"Lais Sampaio de Azevedo , Vanessa Cristina Martins Silva , Raquel Guiducci , Simone Guadagnucci , Fernanda Faria Costa , Monique Beerens Abdul Ghani , Ricardo Duarte Lopes , Antonio Charlys da Costa , Lia Cunha , Marcilio Figueredo Lemos , Adriana Parise , Regina Célia Moreira , Adriana Luchs","doi":"10.1016/j.actatropica.2024.107449","DOIUrl":"10.1016/j.actatropica.2024.107449","url":null,"abstract":"<div><div>Recent increases in zoonotic diseases underscore the integration of companion animals into urban environments, posing complex transmission risks and highlighting the necessity of One Health approaches. Respiratory and enteric viruses have been consistently linked to interspecies transmission between humans and animals. This study aimed to assess the circulation of human noroviruses (NoV), human adenoviruses (HAdV), enteroviruses (EV), parechoviruses (PeV-A), human bocaviruses (HBoV), hepatitis A (HAV) and E viruses (HEV), Influenza A and B viruses (Flu A/B), respiratory syncytial virus (RSV), and SARS-CoV-2 in domestic dogs and cats in Brazil to understand potential zooanthroponosis risks. Between 2012 and 2021, 600 fecal samples from dogs and cats (516 and 84, respectively) were collected at small animal clinics in São Paulo state, Brazil. The specimens underwent in-house qPCR screening for HBoV and HAdV, while EV, PeV-A, NoV, and HEV were tested using in-house RT-qPCR. SARS-CoV-2, Flu A/B, and RSV were investigated with a commercial RT-qPCR kit assay. HAV detection utilized conventional nested (RT)-PCR. Positive samples were sequenced for molecular characterization and phylogenetic analysis. NoV was detected in 0.2 % (1/600) of the animals, while all other investigated viruses tested negative. The NoV-positive sample, collected in 2012 from a pet dog, was identified as genotype GII.4_Sydney[P31]. The Dog/BRA/2012/GII.4_Sydney[P31]/IAL-M21 strain exhibited a close genetic relationship to Brazilian human and environmental NoV GII.4_Sydney[P31] strains, with 98.1–99.2 % nucleotide similarity in ORF1 and 99.2–99.6 % in ORF2 sequences, suggesting interspecies transmission. Pet dogs are frequently exposed to human fecal-borne viruses, highlighting the potential for zooanthroponotic transmission due to their close interaction with humans in shared environments. There is an urgent need to enhance surveillance studies in companion animals to better understand the implications of detecting human NoV strains in pets, as NoV could potentially act as a reverse zoonotic disease in households, animal hospitals, or shelters worldwide.</div></div>","PeriodicalId":7240,"journal":{"name":"Acta tropica","volume":"260 ","pages":"Article 107449"},"PeriodicalIF":2.1,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142556884","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
In recent days, in tropical and subtropical regions, secondary vectors of Anopheles mosquitoes are becoming more important in transmitting diseases to humans as primary vectors. Various molecular techniques have separated closely related Anopheles subpictus and Anopheles vagus mosquitoes based on their diversity with other mosquito species. Despite their widespread distribution, the An. subpictus and An. vagus mosquitoes, which carry Plasmodium in their salivary glands, were not considered primary malaria vectors in India. An. vagus mosquitoes are zoophilic and physically similar to An. subpictus. We intend to identify An. subpictus and An. vagus mosquito's sister species based on their Interspaced Transcribed Region-2 (ITS2). We isolated the midgut gDNA from each mosquito and used ITS2-PCR and Sanger sequencing to characterize the mosquito species. BioEdit software aligned the sequences, and MEGA7 built a phylogenetic tree from them. According to this study, the information gathered from these mosquito samples fits the An. subpictus species A form and the An. vagus Indian form. Furthermore, gut microbiome plays an important role in providing nutrients, immunity, and food processing, whereas mosquitoes' midgut microbiota changes their hosts and spreads illnesses. So, we used the Illumina sequencer to look at the gut microbiome diversity of An. subpictus and An. vagus mosquitoes using 16S rRNA-based metagenomic sequencing. Both mosquito species had an abundant phylum of Pseudomonadota (Proteobacteria), Bacillota, Bacteroidota, and Actinomycetota in their gut microbiomes. Notably, both mosquito species had the genus Serratia in their gut. In the subpictus midgut, the genus of Haematosprillum bacteria was dominant, whereas in the vagus mosquito, the genus of Salmonella was dominant. Notably, current research has observed the Sodalis spp. Bacterial genus for the first time.
近来，在热带和亚热带地区，按蚊的次级传播媒介在向人类传播疾病方面正变得像一级传播媒介一样重要。各种分子技术根据亚按蚊（Anopheles subpictus）和按蚊（Anopheles vagus）与其他蚊子物种的多样性，将这两种密切相关的蚊子区分开来。尽管亚按蚊和迷走蚊分布广泛，但它们的唾液腺中携带疟原虫，在印度并不被认为是疟疾的主要传播媒介。迷走蚊具有嗜动物性，与亚疟蚊体形相似。我们打算根据它们的间距转录区-2（ITS2）来确定亚爪蝇蚊和迷走蚊的姊妹种。我们分离了每只蚊子的中肠 gDNA，并使用 ITS2-PCR 和 Sanger 测序来确定蚊子的种类。BioEdit 软件对序列进行了比对，MEGA7 据此构建了系统发生树。根据这项研究，从这些蚊子样本中收集到的信息符合亚型疟蚊（An. subpictus species A form）和印度型疟蚊（An. vagus Indian form）。此外，肠道微生物群在提供营养、免疫力和食物加工方面发挥着重要作用，而蚊子的中肠微生物群会改变宿主并传播疾病。因此，我们利用Illumina测序仪，使用基于16S rRNA的元基因组测序技术，研究了亚疟蚊和迷走蚊的肠道微生物组多样性。这两种蚊子的肠道微生物组中都有丰富的假单胞菌门（蛋白菌门）、芽孢杆菌门、类杆菌门和放线菌门。值得注意的是，两种蚊子的肠道中都有沙雷氏菌属。在亚目蚊子的中肠中，血吸虫属细菌占主导地位，而在迷走蚊子中，沙门氏菌属细菌占主导地位。值得注意的是，目前的研究首次观察到了索达利属细菌。
{"title":"The genetic composition of Anopheles mosquitoes and the diverse population of gut-microbiota within the Anopheles subpictus and Anopheles vagus mosquitoes in Tamil Nadu, India","authors":"Sathishkumar Vinayagam , Kathirvel Sekar , Devianjana Rajendran , Karthikeyan Meenakshisundaram , Ashish Panigrahi , Dhanush Kumar Arumugam , Ipsita Pal Bhowmick , Kamaraj Sattu","doi":"10.1016/j.actatropica.2024.107439","DOIUrl":"10.1016/j.actatropica.2024.107439","url":null,"abstract":"<div><div>In recent days, in tropical and subtropical regions, secondary vectors of Anopheles mosquitoes are becoming more important in transmitting diseases to humans as primary vectors. Various molecular techniques have separated closely related <em>Anopheles subpictus</em> and <em>Anopheles vagus</em> mosquitoes based on their diversity with other mosquito species. Despite their widespread distribution, the <em>An. subpictus</em> and <em>An. vagus</em> mosquitoes, which carry <em>Plasmodium</em> in their salivary glands, were not considered primary malaria vectors in India. An. vagus mosquitoes are zoophilic and physically similar to An. subpictus. We intend to identify An. subpictus and An. vagus mosquito's sister species based on their Interspaced Transcribed Region-2 (ITS2). We isolated the midgut gDNA from each mosquito and used ITS2-PCR and Sanger sequencing to characterize the mosquito species. BioEdit software aligned the sequences, and MEGA7 built a phylogenetic tree from them. According to this study, the information gathered from these mosquito samples fits the <em>An. subpictus</em> species A form and the <em>An. vagus</em> Indian form. Furthermore, gut microbiome plays an important role in providing nutrients, immunity, and food processing, whereas mosquitoes' midgut microbiota changes their hosts and spreads illnesses. So, we used the Illumina sequencer to look at the gut microbiome diversity of <em>An. subpictus</em> and <em>An. vagus</em> mosquitoes using 16S rRNA-based metagenomic sequencing. Both mosquito species had an abundant phylum of Pseudomonadota (Proteobacteria), Bacillota, Bacteroidota, and Actinomycetota in their gut microbiomes. Notably, both mosquito species had the genus Serratia in their gut. In the <em>subpictus</em> midgut, the genus of Haematosprillum bacteria was dominant, whereas in the <em>vagus</em> mosquito, the genus of <em>Salmonella</em> was dominant. Notably, current research has observed the <em>Sodalis spp</em>. Bacterial genus for the first time.</div></div>","PeriodicalId":7240,"journal":{"name":"Acta tropica","volume":"260 ","pages":"Article 107439"},"PeriodicalIF":2.1,"publicationDate":"2024-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142543040","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-29DOI: 10.1016/j.actatropica.2024.107441
Angelita Fernandes Druzian , Adriana de Oliveira França , Minoru German Higa-Júnior , Maria Elizabeth Cavalheiros Dorval , Manoel Sebastião da Costa Lima-Junior , Mauricio Antonio Pompilio , Maria de Fatima Cepa Matos , Lídia Raquel de Carvalho , Rinaldo Poncio Mendes , Anamaria Mello Miranda Paniago
Visceral leishmaniasis (VL) poses a serious health threat, particularly when untreated, necessitating accurate diagnosis. While the gold-standard method involves identifying amastigotes in bone marrow aspirate (BMA), this procedure is invasive and occasionally contraindicated. Additionally, when VL is associated with HIV infection the serologies accuracies could be affected. This study aims to evaluate and compare diagnostic methods for VL in patients with and without HIV coinfection. We enrolled prospectively 127 consecutive adult VL patients, 48 (37.8%) of whom had HIV coinfection, in Brazil's Midwestern region, where VL is endemic. Parasitological examination served as the reference standard for accuracy analysis, with index tests including immunofluorescent antibody test (IFAT), immunochromatographic test with rK39 protein (rK39-ICT), and blood polymerase chain reaction (PCR). Specificity assessment involved 430 healthy blood donors from the same endemic area. Ninety-two patients had parasitologically confirmed VL. Among HIV-uninfected patients, rK39-ICT exhibited sensitivity comparable to PCR (93.6%; 95% CI: 83.6–100 vs. 97.8%; 95% CI: 93.6–99.2, respectively) and superior to IFAT (71.1%; 95% CI: 57.9–84.3). However, in HIV-infected patients, rK39-ICT sensitivity was notably lower than PCR (40.0%; 95% CI: 22.5–57.5 vs. 97.4%; 95% CI: 92.5–98.9) and similar to IFAT (67.5%; 95% CI: 52.9–82.0). Combining two serological tests in parallel identified 82.1% of parasitologically confirmed VL cases, with a negative likelihood ratio significantly lower than either test alone. No test achieved a specificity of 90%, and there were no significant differences in specificity observed among the index tests. The positivity rate of parasitological examination in the 127 VL patients was higher in HIV-infected compared to HIV-uninfected patients, 91.3% (95% CI: 83.2–99.4) versus 67.6% (95% CI: 56.9–78.3), respectively. These findings underscore the necessity of accounting for HIV infection when choosing VL diagnostic methods. Although rK39-ICT provides reliable results in HIV-uninfected patients, BMA examination remains crucial for accurate diagnosis in individuals with HIV/AIDS. In cases where bone marrow aspiration is contraindicated, employing IFAT and rK39-ICT in parallel could be considered, as the occurrence of both positive results is uncommon in healthy individuals from endemic areas.
内脏利什曼病（VL）对健康构成严重威胁，尤其是在未经治疗的情况下，因此必须进行准确诊断。虽然黄金标准方法是在骨髓抽吸物（BMA）中识别非主凝集体，但这一过程具有侵入性，有时会有禁忌。此外，当 VL 伴有 HIV 感染时，血清学的准确性也会受到影响。本研究旨在评估和比较合并和未合并 HIV 感染患者的 VL 诊断方法。我们在 VL 流行的巴西中西部地区连续招募了 127 名成年 VL 患者，其中 48 人（37.8%）合并有 HIV 感染。寄生虫学检查是准确性分析的参考标准，指标检测包括免疫荧光抗体检测（IFAT）、rK39 蛋白免疫层析检测（rK39-ICT）和血液聚合酶链反应（PCR）。特异性评估涉及来自同一流行地区的 430 名健康献血者。92 名患者经寄生虫学证实患有 VL。在未感染 HIV 的患者中，rK39-ICT 的灵敏度与 PCR 相当（分别为 93.6%；95% CI：83.6-100 与 97.8%；95% CI：93.6-99.2），优于 IFAT（71.1%；95% CI：57.9-84.3）。然而，在 HIV 感染者中，rK39-ICT 的灵敏度明显低于 PCR（40.0%；95% CI：22.5-57.5 vs. 97.4%；95% CI：92.5-98.9），与 IFAT 相似（67.5%；95% CI：52.9-82.0）。同时使用两种血清学检测方法可鉴定出 82.1% 的寄生虫学确诊 VL 病例，其阴性似然比明显低于单独使用任何一种检测方法。没有一种检测的特异性达到 90%，而且各指标检测的特异性没有明显差异。在 127 例 VL 患者中，HIV 感染者的寄生虫学检查阳性率高于 HIV 未感染者，分别为 91.3%（95% CI：83.2-99.4）和 67.6%（95% CI：56.9-78.3）。这些发现强调了在选择 VL 诊断方法时考虑 HIV 感染的必要性。尽管 rK39-ICT 可为未感染 HIV 的患者提供可靠的结果，但骨髓穿刺检查对于准确诊断 HIV 感染者/艾滋病患者仍然至关重要。在骨髓穿刺有禁忌的情况下，可以考虑同时采用 IFAT 和 rK39-ICT 方法，因为在流行地区的健康人中，同时出现这两种阳性结果的情况并不常见。
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