现代畜牧兽医Issue(4)：12-16,5.DOI:10.20154/j.cnki.issn1672-9692.2026.04.003
微小隐孢子虫PCR检测方法的建立与应用
Establishment and application of a PCR assay for Cryptosporidium parvum
摘要
Abstract
This study aimed to establish a PCR method for the detection of Cryptosporidium parvum.Based on the GP60 gene sequence of Cryptosporidium parvum downloaded from GenBank,a pair of specific primers was designed targeting a highly conserved region.The PCR method was established through program optimization,and its sensitivity,specificity,repeatability,and clinical sample detection performance were evaluated.The results showed that the established PCR method could amplify a specific 921 bp fragment of Cryptosporidium parvum,consistent with the expected size.No cross-reactions were observed with enterotoxigenic Escherichia coli K99,Clostridium perfringens,Eimeria sp.,bovine rotavirus,and bovine coronavirus.After serial dilution of positive samples,the minimum detection concentration was 0.243 mg/L.Intra-assay and inter-assay results demonstrated good repeatability and stability.Using this method,411 fecal and anal swab samples from diarrheic cattle collected in Heilongjiang Province were tested,and 141 samples were positive for Cryptosporidium parvum,with an overall positive rate of 34.31%.The study indicates that the established method has good specificity,sensitivity,and repeatability,and is well suited for the detection of clinical samples.关键词
微小隐孢子虫/牛隐孢子虫病/PCR/GP60基因Key words
Cryptosporidium parvum/Bovine crypto-sporidiosis/PCR/GP60 gene分类
农业科技引用本文复制引用
邓河淳,姚冰鑫,杨帆,王俊,韩宇鑫,常继涛,尹鑫..微小隐孢子虫PCR检测方法的建立与应用[J].现代畜牧兽医,2026,(4):12-16,5.基金项目
"十四五"国家重点研发计划"草食家畜重要疫病新型疫苗创制(2023YFD180250300)" （2023YFD180250300）
