用于检测蓝氏贾第鞭毛虫的实时聚合酶链反应

Real-time PCR for the detection of Giardia lamblia.

作者信息

Verweij Jaco J, Schinkel Janke, Laeijendecker Daphne, van Rooyen Marianne A A, van Lieshout Lisette, Polderman Anton M

机构信息

Department of Parasitology and Department of Medical Microbiology, Leiden University Medical Center, P.O. Box 9600, Leiden, The Netherlands.

出版信息

Mol Cell Probes. 2003 Oct;17(5):223-5. doi: 10.1016/s0890-8508(03)00057-4.

DOI:10.1016/s0890-8508(03)00057-4

PMID:14580396

Abstract

Microscopy is considered to be the gold standard for diagnosis of Giardia lamblia infection. However, this method is time-consuming and not very sensitive. We developed a real-time PCR assay based on the small subunit ribosomal RNA gene of G. lamblia for the specific detection of G. lamblia DNA in stool samples and thereafter compared the results with microscopy and antigen detection. The G. lamblia real-time PCR was positive in 102 of 104 fecal samples known to contain G. lamblia cysts and was positive in 10 fecal samples in which G. lamblia antigen was detected but in which no cysts were found with microscopic examination of concentrated fecal samples. The real-time PCR is as specific and sensitive as antigen detection and is more sensitive than microscopy. Moreover, in two patients we were able to detect G. lamblia earlier in the course of infection than with any of the other methods.

摘要

显微镜检查被认为是诊断蓝氏贾第鞭毛虫感染的金标准。然而，这种方法耗时且不太灵敏。我们基于蓝氏贾第鞭毛虫的小亚基核糖体RNA基因开发了一种实时荧光定量聚合酶链反应（PCR）检测方法，用于粪便样本中蓝氏贾第鞭毛虫DNA的特异性检测，随后将结果与显微镜检查和抗原检测进行比较。在已知含有蓝氏贾第鞭毛虫包囊的104份粪便样本中，有102份样本的蓝氏贾第鞭毛虫实时荧光定量PCR检测呈阳性；在10份粪便样本中，蓝氏贾第鞭毛虫抗原检测呈阳性，但浓缩粪便样本的显微镜检查未发现包囊，而这些样本的实时荧光定量PCR检测也呈阳性。实时荧光定量PCR与抗原检测一样特异和灵敏，且比显微镜检查更灵敏。此外，在两名患者中，我们能够比其他任何方法更早地在感染过程中检测到蓝氏贾第鞭毛虫。