Pandey Netranapha, Siripattanapipong Suradej, Leelayoova Saovanee, Manomat Jipada, Mungthin Mathirut, Tan-Ariya Peerapan, Bualert Lertwut, Naaglor Tawee, Siriyasatien Padet, Phumee Atchara, Piyaraj Phunlerd
Department of Microbiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
Department of Parasitology, Phramongkutklao College of Medicine, Bangkok 10400, Thailand.
Acta Trop. 2018 Sep;185:294-300. doi: 10.1016/j.actatropica.2018.06.006. Epub 2018 Jun 8.
Leishmaniasis is a neglected tropical disease causing opportunistic infection among patients with HIV/AIDS. The fatal form of this disease is visceral leishmaniasis (VL). DNA of Leishmania can be detected in saliva, for which the collection is noninvasive and requires little expertise. This study aimed to evaluate the sensitivity and specificity of a nested-PCR to amplify the Internal Transcribed Spacer 1 (ITS1) to detect Leishmania DNA in paired saliva and buffy coat samples of 305 Thai patients with HIV/AIDS in Trang Hospital, Trang Province, southern Thailand. For asymptomatic Leishmania infection among Thai patients with HIV/AIDS, the sensitivity and specificity of the nested-PCR-ITS1 in buffy coat were 73.9 and 100%, respectively. However, the sensitivity in saliva was 26.1% and specificity was 100%. Using the nested-PCR-ITS1, saliva and buffy coat samples showed positive agreement in only 52.0% of patients. Saliva tested results with the nested-PCR-ITS1 showed positive agreement with the Direct Agglutination Test (DAT) in 46.5% of patients. Only 12.1% of the samples showed positive agreement for Leishmania infection among all the three tests: saliva, buffy coat and DAT results. Using nucleotide sequencing, at least three species of Leishmania infection were identified in saliva, i.e., L. siamensis (n = 28), L. martiniquensis (n = 9), and L. donovani complex (n = 1). As a result, buffy coat still appears to be a better specimen to diagnose asymptomatic VL infection among individuals with HIV. However, the use of both buffy coat and saliva together as clinical specimens would increase the sensitivity of Leishmania detection.
利什曼病是一种被忽视的热带疾病，可在艾滋病毒/艾滋病患者中引发机会性感染。该疾病的致命形式是内脏利什曼病（VL）。利什曼原虫的DNA可在唾液中检测到，唾液采集是非侵入性的，且所需专业知识较少。本研究旨在评估巢式聚合酶链反应（nested-PCR）扩增内转录间隔区1（ITS1）以检测泰国南部董里府董里医院305例艾滋病毒/艾滋病患者配对唾液和血沉棕黄层样本中利什曼原虫DNA的敏感性和特异性。对于泰国艾滋病毒/艾滋病患者中的无症状利什曼原虫感染，血沉棕黄层中巢式PCR-ITS1的敏感性和特异性分别为73.9%和100%。然而，唾液中的敏感性为26.1%，特异性为100%。使用巢式PCR-ITS1，唾液和血沉棕黄层样本仅在52.0%的患者中显示出阳性一致性。巢式PCR-ITS1检测的唾液结果与直接凝集试验（DAT）在46.5%的患者中显示出阳性一致性。在唾液、血沉棕黄层和DAT结果这三项检测中，仅12.1%的样本显示出利什曼原虫感染的阳性一致性。通过核苷酸测序，在唾液中鉴定出至少三种利什曼原虫感染，即暹罗利什曼原虫（n = 28）、马提尼克利什曼原虫（n = 9）和杜氏利什曼原虫复合体（n = 1）。因此，血沉棕黄层似乎仍是诊断艾滋病毒感染者无症状VL感染的更好标本。然而，将血沉棕黄层和唾液一起作为临床标本使用将提高利什曼原虫检测的敏感性。
