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基于SYBR Green I染料反应的环介导等温扩增技术用于泌尿生殖系统结核的诊断

Diagnosis of genitourinary tuberculosis by loop-mediated isothermal amplification based on SYBR Green I dye reaction.

作者信息

Kamra Ekta, Singh Netrapal, Khan Anish, Singh Jaideep, Chauhan Meenakshi, Kamal Hemant, Mehta Promod K

机构信息

Centre for Biotechnology, Maharshi Dayanand University, Rohtak, 124001, India.

Noor Medical Centre, Sector-6, Karnal, 132001, India.

出版信息

Biotechniques. 2022 Jun;73(1):47-57. doi: 10.2144/btn-2022-0027. Epub 2022 Jul 5.

Abstract

A multitargeted loop-mediated isothermal amplification (MT-LAMP) assay targeting (Rv1980c) and IS was designed to diagnose genitourinary tuberculosis (GUTB) cases. While assessing gel-based, hydroxynaphthol blue (HNB) and SYBR Green I MT-LAMP assays on GUTB specimens (n = 28) in a pilot study, both gel-based/SYBR Green I assays exhibited better sensitivity than HNB LAMP. Since SYBR Green MT-LAMP is easier to perform compared with a gel-based assay, a higher number of GUTB specimens (n = 55) were evaluated by SYBR Green MT-LAMP, wherein 85.5% sensitivity and 94.4% specificity (n = 36) were obtained. Moreover, the sensitivity attained by MT-LAMP was significantly higher (p < 0.05) than with multiplex-PCR ( + IS). After further validating these MT-LAMP data in different epidemiological settings, this assay may be developed as a diagnostic kit.

摘要

一种针对(Rv1980c)和插入序列(IS)的多靶点环介导等温扩增(MT-LAMP)检测方法被设计用于诊断泌尿生殖系统结核(GUTB)病例。在一项初步研究中,对28份GUTB标本进行基于凝胶的、羟基萘酚蓝(HNB)和SYBR Green I MT-LAMP检测评估时,基于凝胶的/SYBR Green I检测均表现出比HNB LAMP更好的敏感性。由于与基于凝胶的检测相比,SYBR Green MT-LAMP操作更简便,因此通过SYBR Green MT-LAMP对更多数量的GUTB标本(n = 55)进行了评估,其中获得了85.5%的敏感性和94.4%的特异性(n = 36)。此外,MT-LAMP获得的敏感性显著高于多重聚合酶链反应(针对 + IS)(p < 0.05)。在不同的流行病学环境中进一步验证这些MT-LAMP数据后,该检测方法可能会被开发成诊断试剂盒。

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