SYBR GreenⅠ实时荧光定量RT-PCR与常规RT-PCR检测大熊猫轮状病毒的比较研究  

Comparative study of SYBR Green Ⅰreal time RT-PCR and conventional RT-PCR for detecting giant panda rotavirus

作　　者：曾杨茹 颜其贵[1] 杨锐[1] 黄杰[1] 任露[1] 王成东 陈欣 王娅[1] 

机构地区：[1]四川农业大学,四川成都611130 [2]成都大熊猫繁育研究基地,四川成都610081

出　　处：《中国兽医科学》2016年第10期1248-1252,共5页Chinese Veterinary Science

基　　金：成都大熊猫繁育研究基金会项目(CPF研2012-9;CPF研2012-12)

摘　　要：为确定一种准确快速检测大熊猫轮状病毒的方法,同时监测大熊猫轮状病毒的感染情况,通过SYBR GreenⅠ实时荧光定量RT-PCR和常规RT-PCR检测2011至2015年(除2014年)大熊猫基地A和B共227份大熊猫粪便中的轮状病毒。结果显示,利用荧光定量RT-PCR方法检测出25份阳性粪便样品,阳性率为11%(25/227);利用常规RT-PCR检出15份阳性粪便,阳性率为6.6%(15/227),表明荧光定量RT-PCR更适用于大熊猫轮状病毒的检测。在大熊猫基地A,2011—2013年未检出轮状病毒阳性样品,而2015年6月检出10份阳性样品,阳性率为13.5%(10/74),表明该基地的轮状病毒未被净化;在大熊猫基地B,2011—2013年平均阳性率为19%(12/63),2015年阳性率降至12%(3/25),表明近年轮状病毒在大熊猫中带毒情况逐年缓解。To determine an accurate and rapid method for the detection of giant panda rotavirus（RV）,and monitor RV infection in giant panda,we detected RV in the 227 giant pandas＇ feces by SYBR GreenⅠ real time RT-PCR and conventional RT-PCR,which were collected from The Giant Panda Base A and B from2011 to 2015（except 2014）.The results showed that 25 positive samples were detected by SYBR Green Ⅰreal time RT-PCR,and its positive rate was 11%（25/227）.While 15 positive samples were detected by RT-PCR,and its positive rate was only 6.6%（15/227）,indicating that SYBR GreenⅠreal time RT-PCR was more sensitive to detect RV infection in giant panda.RV positive samples were not detected from 2011 to2013 at The Giant Panda Base A,and 10 positive samples were detected in June 2015.Its positive rate was13.5%（10/74）.The results indicated that RV of the base is not purified.On average,the RV positive rate of The Giant Panda Base B was 19%（12/63） from 2011 to 2013,and positive rate dropped to 12%（3/25）in 2015.The results showed that the RV infection has decreased in recent years.

关 键 词：大熊猫 轮状病毒 反转录-聚合酶链反应 荧光定量反转录-聚合酶链反应 

分 类 号：S852.659.4[农业科学—基础兽医学]