Onderstepoort Veterinary Instititute, Agricultural Research Council, Private Bag X05, Onderstepoort 0110, South Africa.
Vet Parasitol. 2011 Feb 10;175(3-4):356-9. doi: 10.1016/j.vetpar.2010.10.038. Epub 2010 Oct 27.
A real-time PCR assay based on TaqMan probe chemistry was developed for the detection of Theileria parva DNA in blood samples. It uses a Theileria genus-specific PCR primer set and a T. parva-specific probe to amplify and hybridize with a species-specific part of the 18S rRNA gene of the parasite. The test was evaluated using positive and negative reference blood samples and shown to be specific for T. parva. Analytical sensitivity was determined by testing a dilution series of T. parva positive blood. It was shown to be able to detect parasitaemia as low as 2 × 10(-6)%. The Taqman assay results were also compared with that obtained with the real-time hybridization probe PCR assay, which is currently employed as the official test for the diagnosis of T. parva infections in buffalo and cattle and was shown to be equally sensitive. A panel of 1164 field samples was screened using both assays and 164 samples tested positive in both tests, indicating a good correlation.
建立了一种基于 TaqMan 探针化学的实时 PCR 检测方法，用于检测血液样本中的小泰勒虫 DNA。它使用了一种小泰勒虫属特异性 PCR 引物组和小泰勒虫特异性探针，以扩增和与寄生虫 18S rRNA 基因的种特异性部分杂交。该检测方法使用阳性和阴性参考血液样本进行了评估，结果表明该检测方法对小泰勒虫具有特异性。通过测试小泰勒虫阳性血液的稀释系列来确定分析灵敏度。结果表明，该方法能够检测到低至 2×10(-6)％的寄生虫血症。还将 Taqman 检测结果与目前用于水牛和牛小泰勒虫感染诊断的实时杂交探针 PCR 检测方法进行了比较，结果表明该方法同样敏感。使用这两种检测方法对 1164 个现场样本进行了筛查，结果表明，有 164 个样本在两种检测方法中均呈阳性，表明相关性良好。
