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印度内脏利什曼病病例早期检测的流行病学定量PCR研究

Quantitative PCR in epidemiology for early detection of visceral leishmaniasis cases in India.

作者信息

Sudarshan Medhavi, Singh Toolika, Singh Abhishek Kumar, Chourasia Ankita, Singh Bhawana, Wilson Mary E, Chakravarty Jaya, Sundar Shyam

机构信息

Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India.

Departments of Internal Medicine and Microbiology, University of Iowa and the VA Medical Center, Iowa City, Iowa, United States of America.

出版信息

PLoS Negl Trop Dis. 2014 Dec 11;8(12):e3366. doi: 10.1371/journal.pntd.0003366. eCollection 2014 Dec.

Abstract

INTRODUCTION

Studies employing serological, DTH or conventional PCR techniques suggest a vast proportion of Leishmania infected individuals living in regions endemic for Visceral Leishmaniasis (VL) remain asymptomatic. This study was designed to assess whether quantitative PCR (qPCR) can be used for detection of asymptomatic or early Leishmania donovani infection and as a predictor of progression to symptomatic disease.

METHODS

The study included 1469 healthy individuals living in endemic region (EHC) including both serology-positive and -negative subjects. TaqMan based qPCR assay was done on peripheral blood of each subject using kDNA specific primers and probes.

RESULTS

A large proportion of EHC 511/1469 (34.78%) showed qPCR positivity and 56 (3.81% of 1469 subjects) had more than 1 calculated parasite genome/ml of blood. However, the number of individuals with parasite load above 5 genomes/ml was only 20 (1.36% of 1469). There was poor agreement between serological testing and qPCR (k = 0.1303), and 42.89% and 31.83% EHC were qPCR positive in seropositive and seronegative groups, respectively. Ten subjects had developed to symptomatic VL after 12 month of their follow up examination, of which eight were initially positive according to qPCR and among these, five had high parasite load.

DISCUSSION

Thus, qPCR can help us to detect significant early parasitaemia, thereby assisting us in recognition of potential progressors to clinical disease. This test could facilitate early intervention, decreased morbidity and mortality, and possibly interruption of disease transmission.

摘要

引言

采用血清学、迟发型超敏反应或传统聚合酶链反应(PCR)技术的研究表明,生活在内脏利什曼病(VL)流行地区的大量利什曼原虫感染个体仍无症状。本研究旨在评估定量PCR(qPCR)是否可用于检测无症状或早期杜氏利什曼原虫感染,并作为疾病进展为有症状疾病的预测指标。

方法

该研究纳入了1469名生活在流行地区的健康个体(EHC),包括血清学阳性和阴性的受试者。使用基于TaqMan的qPCR检测方法,利用kDNA特异性引物和探针,对每个受试者的外周血进行检测。

结果

很大一部分EHC(511/1469,34.78%)显示qPCR呈阳性,56名(占1469名受试者的3.81%)每毫升血液中计算出的寄生虫基因组数超过1个。然而,寄生虫载量高于5个基因组/毫升的个体仅有20名(占1469名的1.36%)。血清学检测与qPCR之间的一致性较差(κ=0.1303),血清阳性组和血清阴性组中分别有42.89%和31.83%的EHC的qPCR呈阳性。10名受试者在随访检查12个月后发展为有症状的VL,其中8名最初qPCR呈阳性,在这些人中,5名寄生虫载量高。

讨论

因此,qPCR可帮助我们检测到显著的早期寄生虫血症,从而有助于我们识别可能发展为临床疾病的个体。该检测可促进早期干预、降低发病率和死亡率,并可能阻断疾病传播。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f167/4263468/a8bce6a404cf/pntd.0003366.g001.jpg

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