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当前位置: 首页 > 文章 > 环尾狐猴源弓形虫的检测与鉴定 野生动物学报 2016,37 (2) 118-125
Position: Home > Articles > Detection and Identification of Toxoplasma from Ring-Tailed Lemur(Lemur catta) Chinese Journal of Wildlife 2016,37 (2) 118-125

环尾狐猴源弓形虫的检测与鉴定

作  者：
单芬;李康信;徐春忠;陈武;彭仕明;李婉萍;李国清
单  位：
广州动物园;上海市野生动物园有限公司;华南农业大学兽医学院
关键词：
环尾狐猴;刚地弓形虫;荧光定量PCR;PCR
摘  要：
为了确定引起广东环尾狐猴(Lemur catta)呼吸困难、厌食和出血性肺炎症状的病原,从2只死亡的环尾狐猴体内无菌采集心肝肺肾组织样本。通过病原形态学观察,鉴定为弓形虫(Toxoplasma sp.)后,另外采集在狐猴笼舍出现的2只老鼠和1只猫4份组织样本。所有样本分别采用荧光定量PCR及一步法PCR技术进行弓形虫的检测与529 bp重复序列分析。以弓形虫B1基因片断为扩增对象的荧光定量PCR检测显示6份样本的CT为25.84~30.29,溶解曲线中均出现明显峰值,核酸定量峰值均大于阳性对照。用一步法PCR自6份样本中均扩增出529 bp目的片段,测序和Blast分析显示,该目的片段与GenBank中登录号为KC607824和DQ779189的刚地弓形虫(Toxoplasma gondii)核苷酸序列相似性最高,均为98.7%;与国际标准强毒虫株RH的核苷酸相似性为98.5%。这些结果表明,荧光定量PCR技术可以用于环尾狐猴弓形虫病的快速诊断,本次环尾狐猴的死亡可能是由于弓形虫的感染引起的,刚地弓形虫可能由野猫与老鼠传播。
译  名：
Detection and Identification of Toxoplasma from Ring-Tailed Lemur(Lemur catta)
作  者：
Shan Fen;Li Kangxin;Xu Chunzhong;Chen Wu;Peng Shiming;Li Wanping;Li Guoqing;Guangzhou Zoo;College of Veterinary Medicine,South China Agricultural University;Shanghai Wild Animal Park Company Limited;
关键词：
Ring-tailed lemur;;Toxoplasma gondii;;Fluorogenic quantitative PCR;;Polymerase Chain Reaction
摘  要：
We sampled tissues of heart,liver,lung and kidney from two dead Ring- tailed lemurs(Lemur catta)to identify the pathogens of lemurs that showed symptoms of dyspnea,anorexia and hemorrhagic pneumonia in Guangdong.After the pathogen was identified by morphological observation as Toxoplasma,4 tissue samples from one cat and two rats which appeared in the enclosure of the dead lemurs were collected.All samples were examined by fluorescent quantitative PCR and one step PCR that were specific for Toxoplasma.We conducted sequence analysis of529 bp fragments that was amplified using one step PCR.The CT values of six samples were25.84-30.29 as determined by the fluorescent quantitative PCR method for detecting the Toxoplasma B1 Gene fragment.There were significant peaks in the dissolution cun/es of six samples whose quantitative nucleic acid peak values were greater than those of a positive control.The 529 bp DNA fragments were amplified and cloned from all six samples by one step PCR.The target DNA fragment was sequenced and analyzed using BLAST in GenBank.The result showed that the fragment had the highest similarity(98.7%) with 2 Toxoplasma gondii sequences(accession numbers:KC607824 and DQ779189)and with the international standard virulent strain of Toxoplasma gondii the RH was 98.5%.Generally,the fluorescent quantitative PCR can be considered a rapid diagnostic technique for detecting toxoplasmosis of ring-tailed lemur.The death of ring-tailed lemur might be caused by Toxoplasma gondii which might be spread by cat and rat.

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