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东方次睾吸虫PCR检测方法的建立及初步应用 被引量:7

Establishment and Application of PCR Methods of Meotrchis orientalis
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摘要 为了快速准确地检测鸭、犬等动物东方次睾吸虫的感染情况,根据Gen Bank中发表的东方次睾吸虫ITS序列设计一对特异性引物,以粪便样品中虫卵DNA为模板,优化PCR反应条件,经过敏感性、特异性和重复性实验,建立检测动物粪便中东方次睾吸虫虫卵的方法。结果,阳性样品可扩增得到260 bp的特异性条带,华支睾吸虫、卷棘口吸虫、纤细背孔吸虫DNA样品检测均为阴性,检测到的最低虫卵数为0.031 25个/克粪便,表明研究建立的PCR检测方法具有较高的特异性和灵敏性。临床检测87份鸭粪样本,其中6份为阳性,阳性率为6.9%,结果显示大庆地区鸭子存在东方次睾吸虫感染,所建立的方法适合于鸭子及其他家禽东方次睾吸虫的检测。 To establish a method for detecting Meotrchis orientalis,two specific primers were synthesized according to the sequence of ITS r DNA of M.orientalis on the Gen Bank to amplify the DNA in the eggs by optimizing PCR conditions,then evaluating sensitivity,specificity and repeatability. Finally,a method of detect ing M.orientalis was set up.The results showed that the obtained specific band was 260 bp,and under the same constraint,PCR assay was negative to Clonorchis sinensis,Echinostoma revolutum,Notocotylus attenuatus,Echinostoma revolutum DNA. The method's minimum detect limit was 0.031 25 eggs DNA per gram feces. Among 87 clinical samples there were six positive samples,so the positive rate was 6.9%. The results indicated that there was M.orientalis in the poultry farm in Daqing city.The PCR method had high sensitivity and specificity,which could be used to diagnose M.orientalis in ducks and other poultry.
出处 《黑龙江八一农垦大学学报》 2016年第3期43-45,117,共4页 journal of heilongjiang bayi agricultural university
基金 黑龙江省青年科学基金项目(QC2014C033) 家畜疫病病原生物学国家重点实验室开放基金(SKLVEB2014KFKT006) 国家大学生创新创业训练计划项目(201410223001)
关键词 东方次睾吸虫 PCR检测方法 建立 应用 Metorchis orientalis PCR method establishment application
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