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如何提高克氏锥虫感染的早期诊断:先天性感染儿童中经验证的传统诊断与定量 DNA 扩增的关系。

How to improve the early diagnosis of Trypanosoma cruzi infection: relationship between validated conventional diagnosis and quantitative DNA amplification in congenitally infected children.

机构信息

Instituto Nacional de Parasitología (INP) Dr. M. Fatala Chaben, Paseo Colón 568 (1063), Administración Nacional de Laboratorios e Institutos de Salud (ANLIS) Buenos Aires, Argentina.

出版信息

PLoS Negl Trop Dis. 2013 Oct 17;7(10):e2476. doi: 10.1371/journal.pntd.0002476. eCollection 2013.

Abstract

BACKGROUND

According to the Chagas congenital transmission guides, the diagnosis of infants, born to Trypanosoma cruzi infected mothers, relies on the detection of parasites by INP micromethod, and/or the persistence of T. cruzi specific antibody titers at 10-12 months of age.

METHODOLOGY AND PRINCIPAL FINDINGS

Parasitemia levels were quantified by PCR in T. cruzi-infected children, grouped according to the results of one-year follow-up diagnosis: A) Neonates that were diagnosed in the first month after delivery by microscopic blood examination (INP micromethod) (n = 19) had a median parasitemia of 1,700 Pe/mL (equivalent amounts of parasite DNA per mL); B) Infants that required a second parasitological diagnosis at six months of age (n = 10) showed a median parasitemia of around 20 Pe/mL and 500 Pe/mL at 1 and 6 months old, respectively, and C) babies with undetectable parasitemia by three blood microscopic observations but diagnosed by specific anti - T. cruzi serology at around 1 year old, (n = 22), exhibited a parasitemia of around 5 Pe/mL, 800 Pe/mL and 20 Pe/mL 1, 6 and 12 month after delivery, respectively. T. cruzi parasites were isolated by hemoculture from 19 congenitally infected children, 18 of which were genotypified as DTU TcV, (former lineage TcIId) and only one as TcI.

SIGNIFICANCE

This report is the first to quantify parasitemia levels in more than 50 children congenitally infected with T. cruzi, at three different diagnostic controls during one-year follow-up after delivery. Our results show that the parasite burden in some children (22 out of 51) is below the detection limit of the INP micromethod. As the current trypanocidal treatment proved to be very effective to cure T. cruzi - infected children, more sensitive parasitological methods should be developed to assure an early T. cruzi congenital diagnosis.

摘要

背景

根据恰加斯先天性传播指南,对于感染了克氏锥虫的母亲所生婴儿的诊断,依赖于通过 INP 微量法检测寄生虫,和/或在 10-12 个月龄时持续存在克氏锥虫特异性抗体滴度。

方法和主要发现

通过 PCR 定量检测感染克氏锥虫的儿童的寄生虫血症水平,根据一年随访诊断结果分组:A)在出生后第一个月通过显微镜血检(INP 微量法)诊断的新生儿(n=19),中位数寄生虫血症为 1700Pe/mL(相当于每毫升寄生虫 DNA 的量);B)需要在 6 个月龄时进行第二次寄生虫学诊断的婴儿(n=10),在 1 个月和 6 个月龄时分别显示中位数寄生虫血症约为 20Pe/mL 和 500Pe/mL;C)通过三次血液显微镜观察无法检测到寄生虫血症但在大约 1 岁时通过特异性抗克氏锥虫血清学诊断的婴儿(n=22),在分娩后 1、6 和 12 个月时分别显示中位数寄生虫血症约为 5Pe/mL、800Pe/mL 和 20Pe/mL。从 19 例先天性感染的儿童中通过血液培养分离出克氏锥虫寄生虫,其中 18 例被基因分型为 DTU TcV(前系 TcIId),只有 1 例为 TcI。

意义

本报告是第一个在超过 50 名先天性感染克氏锥虫的儿童中定量检测寄生虫血症水平的报告,在分娩后一年的随访期间进行了三次不同的诊断控制。我们的结果表明,一些儿童(22 例中有 22 例)的寄生虫负担低于 INP 微量法的检测限。由于目前的杀锥虫治疗对治愈克氏锥虫感染的儿童非常有效,应该开发更敏感的寄生虫学方法以确保早期的克氏锥虫先天性诊断。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bded/3798617/cf91ac919895/pntd.0002476.g001.jpg

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