[1]龚凤平,张 莹,罗蔼剑,等.马红球菌菌落印迹检测方法的建立[J].中国预防兽医学报,2017,(04):281-285.[doi:10.3969/j.issn.1008-0425.2017.04.07]
 GONG Feng-ping,ZHANG Ying,LUO Ai-jian,et al.Establishment of a colony blot for detection of Rhodococcus equi[J].Chinese journal of preventive veterinary medicine,2017,(04):281-285.[doi:10.3969/j.issn.1008-0425.2017.04.07]
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马红球菌菌落印迹检测方法的建立()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2017年04
页码:
281-285
栏目:
出版日期:
2017-04-15

文章信息/Info

Title:
Establishment of a colony blot for detection of Rhodococcus equi
文章编号:
1008-0589(2017)04-0281-05
作者:
 

龚凤平12张 莹1罗蔼剑1张西君1白 洁1吴 腾1鱼海琼3孙凌霜14*

 (1. 华南农业大学 兽医学院,广东 广州 510642;2. 广东海大畜牧兽医研究院,广东 广州 511400;
3. 广东出入境检验检疫局技术中心,广东 广州 510623;4. 广东省兽医临床重大疾病综合防控重点实验室,广东 广州 510642)
Author(s):
 

GONG Feng-ping12 ZHANG Ying1 LUO Ai-jian1 ZHANG Xi-jun1 BAI Jie1 WU Teng1YU Hai-qiong3 SUN Ling-shuang14*

 (1. College of veterinary medicine, South China Agricultural University, Guangzhou 510642, China;
2. Guangdong Haida Institute of Animal Husbandry & Veterinary,  Guangzhou 511400, China;
3. Guangdong entry exit inspection and Quarantine Bureau technical center, Guangzhou 510623, China;
4. Key Laboratory of comprehensive prevention and control of major diseases in Guangdong Province, Guangzhou 510642, China)
关键词:
 马红球菌菌落印记法土壤
Keywords:
Rhodococcus equi  colony blot test  horses  soil
分类号:
S852.61
DOI:
10.3969/j.issn.1008-0425.2017.04.07
文献标志码:
A
摘要:
为建立快速检测环境中马红球菌(R.equi)的方法,本研究以非致病性R.equi ATCC6939以及致病性R.equi毒力相关脂蛋白A (VapA)分别免疫家兔,制备了兔抗非致病性R.equi多抗与兔抗致病性R.equi VapA蛋白多抗,并以其作为检测抗体,通过优化反应条件,建立了R.equi菌落印迹检测方法。结果显示该方法可特异性检测R.equi并可区分致病性与非致病性R.equi;其R.equi检测敏感性高,特异性强,均明显优于R.equi的PCR鉴定法。利用本研究建立的方法对全国部分地区马场土壤样本的初步调查表明R.equi在部分地区马场土壤中普遍存在,然而未检测到致病性R.equi。本实验建立的检测方法可应用于我国马场环境中R.equi的流行病学调查,为我国马病防控平台提供有力的技术支持,并为中国现代育马场的建设提供必要的参考。
  
Abstract:
Rhodococcus equi is a Gram positive facultative intracellular pathogen, which mainly causes subacute or chronic abscessating bronchopneumonia of foals in foals up to 6 months. So far, the study of R.equi in China is rarely seen, due to lack of the standard pathogenic survey method. To establish a rapid assay for R.equi detection, a colony blot assay was developed with the Rabbit anti-avirulence R.equi polyclonal antibody and Rabbit anti-virulence R.equi VapA polyclonal antibody. After optimizing the reaction conditions, the colony blot was able to specifically detect R.equi and efficiently distinguished the virulence from avirulence R.equi. Compared with R.equi PCR method, the colony blot test had the advantages of high sensitivity and specificity. The established assay could be applied to investigate the R.equi epidemiology in the environment.
  

参考文献/References:

[1] Prescott J F. Rhodococcus equi: an animal and human pathogen.[J]. Clin Microbiol Rev, 1991, 4(1): 20-34.
[2]Giguere S, Cohen N D, Chaffin M K, et al. Diagnosis, treatment, control, and prevention of infections caused by Rhodococcus equi in foals [J]. J Vet Inter Med, 2011, 25(6): 1209-1220.
[3]Hillidge C J. Use of erythromycin-rifampin combination in treatment of Rhodococcus equi pneumonia [J]. Vet Microbiol, 1987, 14(3): 337-342.
[4]Giguere S, Cohen N D, Chaffin M K, et al. Rhodococcusequi: clinical manifestations, virulence, and immunity [J]. J Vet Inter Med, 2011, 25(6): 1221-1230.
[5]Kuskie K R, Smith J L, Wang N, et al. Effects of location for collection of air samples on a farm and time of day of sample collection on airborne concentrations of virulent Rhodococcusequi at two horse breeding farms [J]. Am J Vet Res, 2011, 72(1): 73-79.
[6]龚凤平,孙凌霜,余海琼,等. 马红球菌VapA蛋白的表达及其间接ELISA检测方法的建立[J]. 中国预防兽医学报,2016,38(3):235-239.
[7]Woolcock J B, Farmer A M, Mutime M D. Selective medium for Corynebacterium equi isolation [J]. J Clin Microbiol, 1979, 9(5): 640-642.
[8]Ladron N, Fernandez M, Aguero J, et al. Rapid identification of Rhodococcus equi by a PCR assay targeting the choE gene [J]. JClin Microbiol, 2003, 41(7): 3241-3245.
[9]Haites R E, Muscatello G, Begg A P, et al. Prevalence of the virulence-associated gene of Rhodococcus equi in isolates from  infected foals [J]. J Clin Microbiol, 1997, 35(6): 1642-1644.
[10]Muscatello G, Browning G F. Identification and differentiation of avirulent and virulent Rhodococcus equi using selective media and colony blotting DNA hybridization to determine their concentrations in the environment [J]. Vet Microbiol, 2004, 100(1-2): 121-127.
[11]丁壮,Shinji Takai,Hiroo Madarame,等. 中国内蒙自治区马饲育环境土壤中马红球菌的分布(英文) [J]. 中国兽医学报,2008,28(1):40-44.
[12]Takai S. Epidemiology of Rhodococcus equi infections: a review[J]. Vet Microbiol, 1997, 56(3-4): 167-176.
(本文编辑:彭永刚)

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更新日期/Last Update: 2017-04-27