Laboratório de Investigação Médica (LIM-06) do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo, Brazil; Instituto de Medicina Tropical, Faculdade de Medicina da Universidade de São Paulo, Brazil.
Laboratório de Investigação Médica (LIM-06) do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo, Brazil; Instituto de Medicina Tropical, Faculdade de Medicina da Universidade de São Paulo, Brazil.
Exp Parasitol. 2021 Nov;230:108157. doi: 10.1016/j.exppara.2021.108157. Epub 2021 Sep 17.
Strongyloides venezuelensis has been used in different experimental studies, such as those aimed at the evaluation of diagnostic techniques for human strongyloidiasis, mainly the molecular diagnosis. In this study, three regions (genus, 18S and 28S targets) of Strongyloides ribosomal DNA were evaluated for the molecular diagnosis of experimental strongyloidiasis. Rats were infected subcutaneously with 400 or 4000 S. venezuelensis infective larvae (400iL3 and 4000iL3), and kept for 35 days. Fecal samples were collected daily to count eggs per gram of feces (EPG) and to perform the polymerase chain reaction (PCR). Egg count started on the 5th day post-infection (pi) and ended on days 33 and 34 pi, in 400iL3 and 4000iL3 groups, respectively. Based in EPG, fecal samples were selected from days 2, 5, 8, 11, 15, 23 and 35 pi for DNA extraction; PCR (genus, 18S and 28S); and sequencing. The PCR-28S products showed higher values of identity (95-100%) in the database with the Strongyloides sequences. Therefore, it is possible to reinforce the application of PCR-28S in the diagnosis of experimental and human strongyloidiasis.
委内瑞拉钩虫已被用于不同的实验研究，例如评估人类钩虫病的诊断技术，主要是分子诊断。在这项研究中，对钩虫核糖体 DNA 的三个区域（属、18S 和 28S 靶标）进行了评估，用于实验性钩虫病的分子诊断。老鼠经皮感染 400 或 4000 条委内瑞拉钩虫感染性幼虫（400iL3 和 4000iL3），并保持 35 天。每天收集粪便样本以计算每克粪便中的卵数（EPG）并进行聚合酶链反应（PCR）。卵计数从感染后第 5 天（pi）开始，分别在 400iL3 和 4000iL3 组中持续到第 33 天和第 34 天 pi。根据 EPG，从感染后第 2、5、8、11、15、23 和 35 天的粪便样本中提取 DNA 进行 PCR（属、18S 和 28S）；和测序。PCR-28S 产物在数据库中与钩虫序列具有更高的同一性（95-100%）。因此，PCR-28S 可用于诊断实验性和人类钩虫病。
