技术方法

蓝氏贾第鞭毛虫诊断

高正琴，贺争鸣，岳秉飞

(中国食品药品检定研究院，北京100050)

【摘要】目的蓝氏贾第鞭毛虫作为一种重要的人兽共患寄生虫病贾第虫病的病原，其对SPF实验动物质量造成的潜在威胁不容忽视。本研究的目的是建立蓝氏贾第鞭毛虫快速诊断方法，并对17个生产厂家516批2562只SPF实验动物检查结果进行分析。 方法用直接镜检法、快速姬姆萨染色法和多重PCR方法，对蓝氏贾第鞭毛虫进行检测。 结果在SPF实验动物中检出数量众多的蓝氏贾第鞭毛虫滋养体和包囊，鉴定出蓝氏贾第鞭毛虫18S rDNA、β-giardin、TPI和GDH基因。直接镜检法、快速姬姆萨染色法和多重PCR方法均能检出蓝氏贾第鞭毛虫。17个生产厂家516批2562只SPF实验动物蓝氏贾第鞭毛虫阳性检出率22.9%(586/2562)。 结论直接镜检法、快速姬姆萨染色法和多重PCR方法具有高度的敏感性和特异性，可用于蓝氏贾第鞭毛虫的快速诊断。17个生产厂家516批SPF实验动物蓝氏贾第鞭毛虫检查结果未能全部符合规定。

【关键词】蓝氏贾第鞭毛虫；直接镜检；快速姬姆萨染色；多重PCR；SPF实验动物；质量分析

[基金项目]国家科技支撑课题(2013BAK11B01)。

[作者简介]高正琴(1976-)，女，副研究员，研究方向：病原生物学和快检新技术研究。

[通讯作者]贺争鸣(1957-)，男，研究员，研究方向：实验动物微生物学。

【中图分类号】R332【文献标识码】 A

doi:10.3969.j.issn.1671.7856. 2015.001.014

Diagnoses ofGiardialamblia

GAO Zheng-qin，HE Zheng-ming，YUE Bing-fei

(National Institute for Food and Drug Control, Beijing 10050, China)

Abstract【】ObjectiveGiardia lamblia is an important pathogen of zoonosis giardiasis, it poses a potential threat to the quality of SPF (specific pathogen-free) laboratory animals cannot be ignored. The aim of this study is to establish the method of rapid diagnosis of Giardia lamblia, and analyze the test results of 516 batches form 17 manufactures. Methods Direct microscopy, Giemsa-fast staining and multiplex polymerase chain reaction (multiplex PCR) were applied to detect Giardia lamblia.ResultsNumerous of Giardia lamblia trophozoites and cysts were detected in SPF laboratory animals by using direct microscopy and Giemsa-fast staining, and multiplex PCR were performed to identify 18S rDNA, β-giardin, TPI and GDH genes of DNA extracted from these trophozoites and cysts identified Giardia lamblia. Direct microscopy, Giemsa-fast staining, and multiplex PCR methods can be used to detect Giardia lamblia. Of the 2562 SPF laboratory animals studied, 22.9% (586/2562) were positive for Giardia lamblia.ConclusionsDirect microscopy, Giemsa-fast staining, and multiplex PCR were effective techniques with high sensitivity and specificity for rapid diagnosis of Giardia lambliain. It is not satisfactory that the results of Giardia lamblia examination in 516 batches form 17 manufactures failed to meet the requirements 100%.

【Key words】Giardialamblia；Direct microscopy；Giemsa-fast staining；Multiplex polymerase chain reaction；Specific pathogen-free laboratory animal；Quality analysis

蓝氏贾第鞭毛虫(Giardialamblia)是一种呈全球性分布的寄生性肠道原虫，引起以腹泻、吸收障碍和消化不良为主的蓝氏贾第鞭毛虫病(giardiasis，简称贾第虫病)，也是HIV/AIDS合并感染机会原虫。……