摘要: To establish a TaqMan-based real-time RT-PCR method for detection of canine distemper virus,the pair of primers and the TaqMan-based probe were designed according to the M protein genes of canine distemper virus strains.The reactive conditions were optimized to improve the sensitivity and specificity of the method.The specificity and sensitivity of the method were high.Canine distemper virus in the thirty-eight samples had been tested by the TaqMan-based real-time RT-PCR,the common RT-PCR and electron microscopy.The sensitivity of the TaqMan-based real-time RT-PCR was higher than the common RT-PCR and electron microscopy.This suggests that the method may be used in clinical diagnosis,epizootic study and laboratory research.

摘要: 本文采用巢式PCR/RT-PCR方法,对我国10个动物园中无临床症状的圈养小熊猫的71个肛拭子和61个唾液拭子样品,进行犬瘟热病毒(CDV)、犬细小病毒(CPV)、犬冠状病毒(CCV)、犬腺病毒(CAV)和犬疱疹病毒(CHV)的检测,以评估我国圈养小熊猫是否面临这几种病毒的威胁。对阳性PCR结果进行测序分析,并与GenBank上的序列进行比较。结果,在肛拭子样品中检测到3个CPV和6个CCV阳性结果,经测序后,与GenBank上序列的同源性分别达99%和100%。而在唾液拭子样品中没有检测到任何阳性结果,且CDV、CAV和CCV的检测结果均为阴性。从阳性CPV的肛拭子样品中分离到一株细小病毒毒株,表明圈养小熊猫已受到细小病毒和犬冠状病毒的感染,今后应加强这两种病毒的预防工作。本文所采用的PCR方法检测病毒性疾病,能检测到微量的病毒模板,可对小熊猫病毒性感染进行早期诊断。

摘要: 犬蝠和棕果蝠是西双版纳地区较为常见的两个果蝠物种,大多数时间内它们同域分布,共同利用当地许多野生果实。从2004年6月至12月,我们采用拾遗法、粪便分析法以及种子萌发鉴定法并结合雾网采样对西双版纳地区这两种果蝠的食性进行了初步研究。结果发现犬蝠利用11科18种植物的果实,2科2种植物的叶片;而棕果蝠利用9科12种植物的果实,1科1种植物的叶片。研究发现雨季(6~10月)两种果蝠食物类型在本地区重叠程度较高,它们共同利用的植物类型占记录植物类型总数的65%。在干旱季节(11~12月),棕果蝠避开与犬蝠在食物方面的竞争而去别的地方开拓食物资源。

摘要: In order to investigate the incidence of canine adenovirus(CAV) infection, sera from a total of 109 unvaccinated tigers from 6 districts of China were examined for antibodies to canine adenovirus typeⅠ(CAV-1) and canine adenovirus typeⅡ(CAV-2) by hemagglutination inhibition(HI) test. Antibodies to CAV-1 and CAV-2 were prevalent with positivity rates of 28.4% and 10.1%, respectively. The results suggested that tigers can be infected by CAV.

摘要: 首次对犬冠状病毒大熊猫株(CCVGP)核蛋白(N)基因进行了克隆和序列测定。本实验根据GenBank中报道的CCVlnsavc-1株N蛋白基因序列,设计了一对特异性引物,对分离的CCVGP野毒株进行了RT-PCR扩增。将扩增的PCR产物纯化回收后与pGEM T连接得到重组质粒pTN,进行核苷酸序列测定。结果该基因全长1146bp,编码382个氨基酸;与CCⅤ标准毒株Insavc-lN基因相比,核苷酸的同源性为92 6%,推导的氨基酸的同源性为93 2%。在推导的N蛋白N端156-179位存在一个SRXX富集区,与小鼠肝炎病毒N蛋白相应区域相同,推测可能是RNA结合区。预测的GP株N蛋白疏水性和抗原表位与Insavc-l株N蛋白存在细微的差异。将pTN双酶切,回收目的基因片段克隆到大肠杆菌表达载体pET28a中构建了重组质粒pETN,转化大肠杆茵BL21,用IPTG进行了诱导表达。结果重组菌菌体裂解物经SDS-PAGE电泳可检测到相对分子量为48KD的重组蛋白,免疫印迹法证实该重组蛋白可以与CCV多克隆抗体发生特异性反应。经凝胶薄层扫描分析,重组N蛋白表达量可占菌体蛋白的49 3%,表达的蛋白纯化后可用于建立检测大熊猫CCV抗体间接ELISA用的包被抗原。