Duplex PCR Assay for Simultaneous Detections of <i>Staphylococcus aureus</i> and <i>Bordetella bronchiseptica</i> in Rabbits

WANG Jin-xiang; SANG Lei; SUN Shi-kun; CHEN Dong-jin; CHEN Yan-feng; XIE Xi-ping

doi:10.19303/j.issn.1008-0384.2018.08.002

Volume 33 Issue 8

Mar. 2019

Turn off MathJax

Article Contents

Article Navigation > Fujian Journal of Agricultural Sciences > 2018 > 33(8): 777-780

WANG Jin-xiang, SANG Lei, SUN Shi-kun, CHEN Dong-jin, CHEN Yan-feng, XIE Xi-ping. Duplex PCR Assay for Simultaneous Detections of Staphylococcus aureus and Bordetella bronchiseptica in Rabbits[J]. Fujian Journal of Agricultural Sciences, 2018, 33(8): 777-780. doi: 10.19303/j.issn.1008-0384.2018.08.002

Citation:

WANG Jin-xiang, SANG Lei, SUN Shi-kun, CHEN Dong-jin, CHEN Yan-feng, XIE Xi-ping. Duplex PCR Assay for Simultaneous Detections of Staphylococcus aureus and Bordetella bronchiseptica in Rabbits[J]. Fujian Journal of Agricultural Sciences, 2018, 33(8): 777-780. doi: 10.19303/j.issn.1008-0384.2018.08.002

Citation:

PDF( 1158 KB)

Duplex PCR Assay for Simultaneous Detections of Staphylococcus aureus and Bordetella bronchiseptica in Rabbits

doi: 10.19303/j.issn.1008-0384.2018.08.002

Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture Sciences, Fuzhou, Fujian 350013, China

Received Date: 2018-06-04
Rev Recd Date: 2018-08-01
Publish Date: 2018-08-01

Abstract

Abstract

To develop a duplex PCR assay for simultaneous detections of Staphylococcus aureus and Bordetella bronchiseptica in rabbits, two pairs of specific primers were designed based on the conserved regions of nuc and fimN genes of the pathogens. The methodology with optimized reaction conditions was specific and free of cross-reactions with Pasteurella multocida, Escherichia coli and Clostridieum welchii. The detection limit of the method was 5 pg genome DNA on both pathogens. Moreover, the results obtained by using the duplex PCR assay were in perfect agreement with those determined separately by single PCR determinations. Thus, the newly developed highly specific, sensitive and accurate methodology could be a rapid assay applied for simultaneous identification of S. aureus and B. bronchiseptica present in a diseased rabbit.
- rabbit,
- Staphylococcus aureus,
- Bordetella bronchiseptica,
- duplex PCR assay

FullText(HTML)

References(14)

References

[1]	王孝友, 王永康, 杨睿, 等.兔葡萄球菌的分离与鉴定[J].中国养兔, 2011(4):12-14. doi: 10.3969/j.issn.1005-6327.2011.04.004
[2]	PATHAK A K, BOAG B, POSS M, et al. Seasonal breeding drives the incidence of a chronic bacterial infection in a free-living herbivore population[J]. Epidemiol infect, 2011, 139(8):2110-1219. http://europepmc.org/abstract/MED/20943004
[3]	张亚黎, 孙志强.家兔葡萄球菌病的初步诊断及防治[J].畜牧兽医杂志, 2015, 34(1):132-134. doi: 10.3969/j.issn.1004-6704.2015.01.058
[4]	姜常青, 赵永峰, 刁非非, 等.兔源金黄色葡萄球菌的分离鉴定[J].山东畜牧兽医, 2015, 36(9):10-11. doi: 10.3969/j.issn.1007-1733.2015.09.005
[5]	刘燕, 韦强, 肖琛闻, 等.兔波氏杆菌研究进展及防控对策思考[J].中国养兔, 2014(4):36-38. doi: 10.3969/j.issn.1005-6327.2014.04.010
[6]	王晓芳, 刘燕, 肖琛闻, 等.兔支气管败血波氏杆菌的分离鉴定与耐药基因检测[J].中国畜牧兽医, 2015, 42(3):544-548. http://www.cnki.com.cn/Article/CJFDTOTAL-GWXK201503006.htm
[7]	刘燕, 章春桃, 韦强, 等.兔波氏杆菌fimN蛋白间接ELISA抗体检测方法的建立[J].中国预防兽医学报, 2013, 35(11):899-902. doi: 10.3969/j.issn.1008-0589.2013.11.09
[8]	马超锋, 陈铭, 徐永杰. PCR检测兔支气管败血波氏杆菌方法的建立及应用[J].河南农业科学, 2017, 46(4):124-127. http://d.old.wanfangdata.com.cn/Periodical/hnnykx201704025
[9]	邓钊宾, 余滔, 耿毅, 等.兔源金黄色葡萄球菌的耐药性及耐药基因分析[J].中国预防兽医学报, 2016, 38(1):45-48. doi: 10.3969/j.issn.1008-0589.2016.01.11
[10]	BRAKSTAD O G, AASBAKK K, MAELAND J A. Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene[J]. J Clin Microbiol, 1992, 30(7):1654-1660. http://d.old.wanfangdata.com.cn/OAPaper/oai_pubmedcentral.nih.gov_265359
[11]	向正刚, 耿毅, 张雨薇, 等.四川兔源金黄色葡萄球菌毒力检测及分子分型[J].华南农业大学学报, 2017, 38(4):62-68. http://d.old.wanfangdata.com.cn/Periodical/hnnydxxb201704011
[12]	刘燕, 章春桃, 韦强, 等.兔支气管败血波氏杆菌PCR快速检测方法的建立及应用[J].中国养兔, 2012(4):13-15. doi: 10.3969/j.issn.1005-6327.2012.04.003
[13]	章春桃, 刘燕, 韦强, 等.兔支气管败血波氏杆菌fimN基因的原核表达[J].浙江农业学报, 2011, 23(5):890-893. doi: 10.3969/j.issn.1004-1524.2011.05.006
[14]	凤晓博, 赵青, 林童.食品中金黄色葡萄球菌PCR检测方法的建立[J].青海畜牧兽医杂志, 2014, 44(5):15-16. doi: 10.3969/j.issn.1003-7950.2014.05.009