利用高分辨率熔解曲线分析鉴别牛乳中的某些支原体属和莱氏无胆甾原体。
Discrimination between some Mycoplasma spp. and Acholeplasma laidlawii in bovine milk using high resolution melting curve analysis.
作者信息
Al-Farha Abd Al-Bar, Petrovski Kiro, Jozani Razi, Hoare Andrew, Hemmatzadeh Farhid
机构信息
School of Animal and Veterinary Sciences, Roseworthy Campus, The University of Adelaide, Roseworthy, South Australia, 5371, Australia.
Mosul Technical Institute, Technical Foundation, Northern Technical University, Mosul, 41000, Iraq.
出版信息
BMC Res Notes. 2018 Feb 7;11(1):107. doi: 10.1186/s13104-018-3223-y.
OBJECTIVES
This study aimed to provide a rapid, accurate and cost-effective diagnostic real time polymerase chain reaction-high resolution melting curve assay (PCR-HRM) to identify and distinguish between four different mycoplasmas and Acholeplasma laidlawii isolated at cow-level from a single commercial dairy farm in South Australia. One set of genus-level universal primers was designed targeting the 16S ribosomal RNA gene.
RESULTS
Real time PCR-HRM analysis was able to identify and distinguish between five different mollicutes, namely A. laidlawii, M. arginini, M. bovirhinis, M. bovis and uncultured Mycoplasma. Results were confirmed through sequencing. Our developed assay provides rapid and accurate screening for Mycoplasma mastitis detection.
目的
本研究旨在提供一种快速、准确且经济高效的诊断实时聚合酶链反应-高分辨率熔解曲线分析方法（PCR-HRM），以鉴定和区分从南澳大利亚一个商业奶牛场的奶牛中分离出的四种不同支原体和莱氏无胆甾原体。设计了一组针对16S核糖体RNA基因的属水平通用引物。
结果
实时PCR-HRM分析能够鉴定和区分五种不同的柔膜菌纲微生物，即莱氏无胆甾原体、精氨酸支原体、牛鼻支原体、牛支原体和未培养的支原体。结果通过测序得到证实。我们开发的检测方法为支原体乳腺炎检测提供了快速准确的筛查。
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