Joon Deepali, Nimesh Manoj, Varma-Basil Mandira, Saluja Daman
Dr BR Ambedkar Center for Biomedical Research, University of Delhi, Delhi 110 007, India.
Shri Guru Tegh Bahadur Khalsa College, University of Delhi, Delhi 110 007, India.
J Microbiol Methods. 2017 Aug;139:87-91. doi: 10.1016/j.mimet.2017.05.007. Epub 2017 May 14.
In the present study, IS6110 loop mediated isothermal amplification (LAMP) assay was modified using dUTP-UNG (uracil-DNA N-glycosylase) strategy to prevent carryover contamination, and was evaluated using clinical specimens. The clinical specimens were collected from 236 suspected patients of pulmonary tuberculosis and 315 specimens of suspected patients of extra pulmonary tuberculosis. DNA was extracted from specimens and used as template for nucleic acid amplification. The results were evaluated with culture method as gold standard. Modified IS6110 LAMP assay showed high sensitivity (94.4%) and specificity (97.2%) in specimens collected from suspected pulmonary tuberculosis patients. Sensitivity was comparatively less (86.67%) in extra pulmonary specimens while specificity was 94.04%. In conclusion, IS6110 LAMP assay was modified to prevent carry over contamination and it was validated to be rapid, sensitive and specific method with prospective application in resource-limited settings.
在本研究中，采用dUTP-UNG（尿嘧啶-DNA糖基化酶）策略对IS6110环介导等温扩增（LAMP）检测法进行了改进，以防止污染残留，并使用临床标本进行了评估。临床标本取自236例疑似肺结核患者和315例肺外结核疑似患者的标本。从标本中提取DNA，并用作核酸扩增的模板。以培养法作为金标准对结果进行评估。改良的IS6110 LAMP检测法在疑似肺结核患者的标本中显示出高灵敏度（94.4%）和特异性（97.2%）。肺外标本的灵敏度相对较低（86.67%），而特异性为94.04%。总之，对IS6110 LAMP检测法进行了改进以防止污染残留，并且验证其为一种快速、灵敏且特异的方法，有望在资源有限的环境中应用。
