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用于检测水牛泰勒虫病的终点可视化环介导等温扩增(LAMP)

An endpoint visualization loop-mediated isothermal amplification (LAMP) for detecting bubaline theileriosis.

作者信息

Kumar Sanjeev, Paliwal Sanjhi, Sudan Vikrant, Shanker Daya, Singh Shanker Kumar

机构信息

Department of Parasitology, U. P. Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura, 281001 India.

College of Biotechnology, U. P. Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura, 281001 India.

出版信息

Beni Suef Univ J Basic Appl Sci. 2022;11(1):67. doi: 10.1186/s43088-022-00251-x. Epub 2022 May 13.

Abstract

BACKGROUND

Tropical theileriosis is a significant disease affecting the health and production levels of buffaloes in India. It is caused by an apicomplexan- The timely and accurate detection of infection is vital for implementing a mass vaccination or control programme in a given area under outbreak. Most of the literature concerned with diagnosis of theileriosis revolves around cattle, and practically, there are very limited assays available for detecting bubaline theileriosis. Loop-mediated isothermal amplification (LAMP) assay certainly amplifies the targeted deoxyribosenucleic acid (DNA) with a comparatively higher efficacy, rapidity and sensitivity. Alongside, minimal use of sophisticated instruments in performing LAMP assay is certainly an add on. The present study describes the application of LAMP assay in diagnosing tropical theileriosis in buffaloes alongside, its comparison with polymerase chain reaction (PCR) and blood microscopical examination.

RESULTS

No cross-reaction was seen with DNA of other haemoprotozoan. LAMP was compared with blood microscopy and PCR. LAMP detected infection in 27 out of 100 buffaloes, while blood microscopy and PCR detected disease in 16 and 24 buffaloes, respectively.

CONCLUSION

The sensitivity, specificity and kappa value prediction of LAMP were found to be much higher than the PCR and blood microscopy. The present communication reports the first use of LAMP in detecting theileriosis in buffaloes in the world.

摘要

背景

热带泰勒虫病是一种影响印度水牛健康和生产水平的重要疾病。它由一种顶复门原虫引起。在疫情爆发地区,及时准确地检测感染对于实施大规模疫苗接种或控制计划至关重要。大多数关于泰勒虫病诊断的文献都围绕牛展开,实际上,可用于检测水牛泰勒虫病的检测方法非常有限。环介导等温扩增(LAMP)检测法确实能以相对更高的效率、速度和灵敏度扩增目标脱氧核糖核酸(DNA)。此外,进行LAMP检测时对精密仪器的使用最少,这无疑是一个优点。本研究描述了LAMP检测法在诊断水牛热带泰勒虫病中的应用,并将其与聚合酶链反应(PCR)和血液显微镜检查进行了比较。

结果

未观察到与其他血原虫DNA的交叉反应。将LAMP与血液显微镜检查和PCR进行了比较。LAMP在100头水牛中检测到27头感染,而血液显微镜检查和PCR分别在16头和24头水牛中检测到疾病。

结论

发现LAMP的敏感性、特异性和kappa值预测均远高于PCR和血液显微镜检查。本通讯报道了世界上首次使用LAMP检测水牛泰勒虫病。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/94bb/9099057/c1d52465efb7/43088_2022_251_Fig1_HTML.jpg

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