Bairami Amir, Rezaei Sasan, Rezaeian Mostafa
Dept. of Medical Parasitology and Mycology, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran.
Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Iran J Parasitol. 2016 Jan-Mar;11(1):19-23.
Among the most important parasitic disease, causing diarrhea, Giardia lamblia is noteworthy. Nowadays detection methods for these parasites include parasitological methods such as microscopic examination. The sensitivity of these methods relies on the expertise and experience of examiners. In contrast, molecular methods such as PCR are less dependent on the expertise of the examiner. Here we developed a PCR for the detection of G. lamblia genome in stool samples in comparison with microscopy, which is the gold standard.
For the evaluation of primers, 22 positive samples and 47 negative samples were used. QIAamp DNA Stool Mini Kit (QIAGEN, Germany) was used for DNA extraction from feces. Primers for PCR were designed using Primer-BLAST which uses Primer 3 to designing specific primers (NCBI/Primer-BLAST).
Sensitivity of the PCR was done with 100% (95%CI: 84.56-100) for the detection of G. lamblia DNA isolated from patients stool samples which were positive for G. lamblia cysts and/or trophozoites using microscopy as gold standard. In comparison with microscopy, PCR had showed the specificity of 97.87% (95%CI: 88.71-99.95).
We designed new primers for the Giardia, and PCR method for the rapid and accurate identification of Giardia parasites established. With consideration to the routine diagnosis techniques in medical parasitology and their limitations such as time consuming, laborious, less sensitivity etc. This G. lamblia PCR is a sensitive and specific application for the diagnosis of G. lamblia and provides us a reliable method in the routine intestinal parasitic infection laboratory diagnosis.
在引起腹泻的最重要的寄生虫病中,蓝氏贾第鞭毛虫值得关注。如今,这些寄生虫的检测方法包括显微镜检查等寄生虫学方法。这些方法的敏感性依赖于检查人员的专业知识和经验。相比之下,PCR等分子方法对检查人员专业知识的依赖较小。在此,我们开发了一种用于检测粪便样本中蓝氏贾第鞭毛虫基因组的PCR方法,并与作为金标准的显微镜检查进行比较。
为评估引物,使用了22份阳性样本和47份阴性样本。采用QIAamp DNA粪便迷你试剂盒(德国QIAGEN公司)从粪便中提取DNA。使用Primer-BLAST设计PCR引物,该软件利用Primer 3设计特异性引物(NCBI/Primer-BLAST)。
以显微镜检查作为金标准,对于从粪便样本中分离出的蓝氏贾第鞭毛虫DNA的检测,PCR的敏感性为100%(95%CI:84.56 - 100),这些粪便样本经显微镜检查发现蓝氏贾第鞭毛虫囊肿和/或滋养体呈阳性。与显微镜检查相比,PCR显示出97.87%(95%CI:88.71 - 99.95)的特异性。
我们设计了针对贾第虫的新引物,并建立了用于快速准确鉴定贾第虫寄生虫的PCR方法。考虑到医学寄生虫学中的常规诊断技术及其局限性,如耗时、费力、敏感性较低等。这种蓝氏贾第鞭毛虫PCR是诊断蓝氏贾第鞭毛虫的一种敏感且特异的应用方法,为我们在常规肠道寄生虫感染实验室诊断中提供了一种可靠的方法。