Laboratorio de Biología Molecular de la Enfermedad de Chagas, Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI-CONICET), Ciudad Autónoma de Buenos Aires, Argentina.
Trans R Soc Trop Med Hyg. 2011 Oct;105(10):543-9. doi: 10.1016/j.trstmh.2011.07.003. Epub 2011 Jul 30.
Chagas disease ranks among the world's most neglected tropical diseases and congenital transmission is increasingly responsible for urbanization of Chagas disease in non-endemic areas. Molecular assays for amplification and profiling of parasite minicircle DNA (kDNA) and identification of discrete typing units (DTUs) were prospectively conducted in bloodstream and placental samples from pregnant women cursing chronic Chagas disease residing in Buenos Aires city. Sensitivity of kDNA-PCR increased from 75.6% to 95.6% when one to three sequential blood samples were analysed. Congenital infection (CI) was diagnosed in 3 neonates born to kDNA-PCR positive mothers, one who had transmitted CI in a previous gestation, pointing to family clustering of congenital transmission. Fourteen of 44 placental samples were kDNA-PCR positive, all from non-CI transmitting women, indicating that placental PCR is not useful for CI diagnosis. Placental PCR positivity was not related to maternal bloodstream PCR positivity and placental parasitic subpopulations not observed in bloodstream were detected by minicircle signatures. PCR targeted to intergenic regions of spliced-leader genes and serological tests using trypomastigote small surface recombinant antigens showed predominance of DTU group TcII/V/VI and only one patient infected with TcI. To our knowledge, this is the first PCR-based follow-up study of bloodstream and placental T. cruzi infections during pregnancy, including identification of DTUs. kDNA-PCR assays in serial blood samples provided high sensitivity for detection of T. cruzi DNA in pregnant women with chronic Chagas disease.
恰加斯病属于世界上最被忽视的热带病之一,先天性传播越来越成为非流行地区恰加斯病城市化的原因。本研究前瞻性地对来自布宜诺斯艾利斯市患有慢性恰加斯病的孕妇的血流和胎盘样本进行了寄生虫微小环 DNA(kDNA)扩增和分析以及离散型单位(DTU)鉴定的分子检测。当分析一到三个连续的血样时,kDNA-PCR 的敏感性从 75.6%增加到 95.6%。在 3 名来自 kDNA-PCR 阳性母亲的新生儿中诊断出先天性感染(CI),其中 1 名母亲在之前的妊娠中传播了 CI,这表明先天性传播存在家庭聚集现象。44 个胎盘样本中有 14 个为 kDNA-PCR 阳性,均来自非 CI 传播的女性,这表明胎盘 PCR 对 CI 诊断没有用。胎盘 PCR 阳性与母亲血流 PCR 阳性无关,并且通过微小环特征检测到在血流中未观察到的胎盘寄生虫亚群。针对剪接引导 RNA 基因的间区的 PCR 和使用锥虫小表面重组抗原的血清学检测显示,DTU 组 TcII/V/VI 占主导地位,只有 1 名患者感染了 TcI。据我们所知,这是首次在怀孕期间对血流和胎盘 T. cruzi 感染进行基于 PCR 的随访研究,包括鉴定 DTU。在患有慢性恰加斯病的孕妇中,连续血样的 kDNA-PCR 检测对 T. cruzi DNA 的检测具有很高的敏感性。