南方农业学报2012，Vol.43Issue(5)：701-704,4.DOI:10.3969/j:issn.2095-1191.2012.05.701
安氏隐孢子虫ITS-1基因PCR检测方法的建立
Establishment of PCR assay for detection of ITS-1 gene in Cryptosporidium andersoni
摘要
Abstract
[ Objective ]The aim of this research was to develop an swiftly efficient PCR detection assay for Cryp-tosporidium andersoni to provide technical support for investigation of molecular epidemiology. [Method]A pair of specific primer was designed using published ITS-1 gene sequence and a PCR assay system was developed to conduct specificity test, sensitivity test, clinical detection and conventional nest PCR. [Result]The results showed that the specific band was only amplified from C. andersoni using this PCR assay, but not amplified from C. suis, C. bailey, C. parvum, Theileria sergenti, Babesia bovis, Trypanosoma evansi and Oesophagostomum. The BLAST comparative result revealed that these sequences had above 99% homology with the some sequences published in GenBank. The sensitivity of the assay was 5 oocysts per gram in cattle feces. Total 1613 dairy fecal samples were collected from different areas of Guangxi and tested by the newly developed PCR assay and conventional nest PCR, for which the positive detection rate was both recorded as 11.72%. It was proved that the efficiency of these two methods was 100%. [Conclusion]The established PCR assay was suitable for detection of ITS-1 gene in C. andersoni and molecular epidemics investigation because of its rapidness, sensitivity and specificity.关键词
安氏隐孢子虫/ITS-1基因/PCR检测/巢式PCRKey words
Cryptosporidium andersoni/ ITS-1 gene/ PCR detection/ nest PCR分类
农业科技引用本文复制引用
彭昊,许力干,谢永平,杨威,李军,陶立,陈泽祥,唐林生,魏晓瑞,马春霞,禤雄标,胡帅..安氏隐孢子虫ITS-1基因PCR检测方法的建立[J].南方农业学报,2012,43(5):701-704,4.基金项目
公益性行业(农业)科研专项项目(201103008) （农业）
广西科研基本业务费专项项目(桂科专项12-2) （桂科专项12-2）
广西水产畜牧兽医局资助项目(桂渔牧财[2011]52号) （桂渔牧财[2011]52号）
