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实时聚合酶链反应:检测轻度和无症状贾第虫感染的优势。

Real-time PCR: Benefits for Detection of Mild and Asymptomatic Giardia Infections.

作者信息

Prasertbun Rapeepun, Sukthana Yaowalark, Popruk Supaluk

机构信息

Department of Protozoology, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand.

出版信息

Trop Med Health. 2012 Jun;40(2):31-5. doi: 10.2149/tmh.2012-08. Epub 2012 Jun 27.

Abstract

The majority of Giardia infections are transmitted by the fecal-oral route and cause giardiasis. Children who live in crowded conditions or low socio-economic areas are the risk group for Giardia infection. Interestingly, most of them are asymptomatic or only mildly infected and can shed the Giardia cysts in the environment. Thus, the diagnosis of Giardia infection in asymptomatic or mild infection plays an important role in achieving control of Giardia duodenalis transmission. The objective of this study was to examine parasitic infections using microscopy and to develop a real-time PCR method for detection of Giardia infection in the stool samples of children living on the Thai-Myanmar border. Both species-specific primers and fluorescent labeled G. duodenalis probe were designed using small-subunit ribosomal RNA (ssrRNA). The results showed that 10 (7.69%) and 40 (30.77%) of 130 stool samples were positive for G. duodenalis by microscopy and real-time PCR respectively. Only 3 out of 9 liquid stools revealed G. duodenalis positive using microscopy, but all of them were G. duodenalis-positive using real-time PCR. The detection limit of real-time PCR for G. duodenalis was 0.1 pg/25 µl reaction. It can detect both mild and asymptomatic Giardia infections in children living on the Thai-Myanmar border.

摘要

大多数贾第虫感染通过粪口途径传播并导致贾第虫病。生活在拥挤环境或社会经济地位较低地区的儿童是贾第虫感染的风险群体。有趣的是,他们中的大多数无症状或仅轻度感染,并且可以在环境中排出贾第虫囊肿。因此,在无症状或轻度感染中诊断贾第虫感染对于实现对十二指肠贾第虫传播的控制起着重要作用。本研究的目的是使用显微镜检查寄生虫感染,并开发一种实时PCR方法来检测生活在泰缅边境的儿童粪便样本中的贾第虫感染。使用小亚基核糖体RNA(ssrRNA)设计了物种特异性引物和荧光标记的十二指肠贾第虫探针。结果显示,130份粪便样本中分别有10份(7.69%)和40份(30.77%)通过显微镜检查和实时PCR检测出十二指肠贾第虫阳性。9份液体粪便中只有3份通过显微镜检查显示十二指肠贾第虫阳性,但使用实时PCR检测时所有样本均为十二指肠贾第虫阳性。十二指肠贾第虫实时PCR的检测限为0.1 pg/25 μl反应。它可以检测泰缅边境地区儿童的轻度和无症状贾第虫感染。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7124/3475312/855aa3394a9f/tmh-2012-08-g001.jpg

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