Siriyasatien Padet, Chusri Sarunyou, Kraivichian Kanyarat, Jariyapan Narissara, Hortiwakul Thanaporn, Silpapojakul Khachornsakdi, Pym Adam M, Phumee Atchara
Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
Excellence Center for Emerging Infectious Disease, King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, 10330, Thailand.
BMC Infect Dis. 2016 Feb 24;16:89. doi: 10.1186/s12879-016-1433-2.
Leishmaniasis caused by two new species of Leishmania; L. siamensis and L. martiniquensis have been recently described in Thailand. The disease has mainly been documented in AIDS patients from southern Thailand. In this study, polymerase chain reaction (PCR) was used to determine HIV-Leishmania co-infection in southern Thailand.
One ml of saliva and 3 ml of EDTA blood were collected from HIV-infected patients for PCR detection of Leishmania DNA, cloning and sequencing. The positive PCR samples were then cultured on Schneider's insect medium.
Three out of 316 saliva samples collected from HIV-infected patients were found to be positive for Leishmania DNA (0.95%). Among the positive samples, one patient was observed with disseminated cutaneous lesions and also tested positive via saliva, whole blood and buffy coat in PCR. The second case presenting with nodular lesions also gave a positive saliva test via PCR two months prior to buffy coat. This diagnosis was confirmed by microscopic examination and a culture of biopsy samples from a nodule. The last case was an asymptomatic Leishmania infection which tested PCR positive only in saliva with a consecutive sample collection conducted for three months.
The prevalence of Leishmania infection in HIV infected patients within this study is 0.95%. Leishmania DNA was detected in saliva by PCR prior to blood and buffy coat of two HIV infected patients. Early detection of Leishmania DNA in saliva would be beneficial for the follow up of asymptomatic Leishmania infected patients, the early treatment of leishmaniasis and for surveillance survey purpose. However, full evaluation of sensitivity and specificity of this technique with a large cohort of patients is required before deployment.
最近在泰国发现由两种新型利什曼原虫,即暹罗利什曼原虫和马提尼克利什曼原虫引起的利什曼病。该疾病主要在泰国南部的艾滋病患者中被记录。在本研究中,采用聚合酶链反应(PCR)来确定泰国南部艾滋病毒与利什曼原虫的合并感染情况。
从感染艾滋病毒的患者中采集1毫升唾液和3毫升乙二胺四乙酸(EDTA)抗凝血,用于PCR检测利什曼原虫DNA、克隆及测序。然后将PCR阳性样本接种于施奈德昆虫培养基上培养。
从感染艾滋病毒的患者中采集的316份唾液样本中,有3份利什曼原虫DNA检测呈阳性(0.95%)。在阳性样本中,一名患者出现播散性皮肤病变,其唾液、全血和血沉棕黄层的PCR检测均呈阳性。第二例表现为结节性病变的患者,在血沉棕黄层检测阳性前两个月,唾液PCR检测就呈阳性。通过显微镜检查和对结节活检样本进行培养,确诊了该病例。最后一例是无症状利什曼原虫感染,仅唾液PCR检测呈阳性,连续三个月采集样本均为此结果。
本研究中感染艾滋病毒患者的利什曼原虫感染率为0.95%。两名感染艾滋病毒的患者在血液和血沉棕黄层检测之前,通过PCR在唾液中检测到了利什曼原虫DNA。唾液中利什曼原虫DNA的早期检测,将有利于无症状利什曼原虫感染患者的随访、利什曼病的早期治疗以及监测调查。然而,在推广应用之前,需要对大量患者进行该技术的敏感性和特异性的全面评估。