Diagnosis of pleural tuberculosis by multi-targeted loop-mediated isothermal amplification assay based on SYBR Green I reaction: comparison with GeneXpert® MTB/RIF assay

被引：2

作者：

Soni, Aishwarya ^{[1
,2
]}

Dahiya, Bhawna ^{[1
,3
]}

Sheoran, Abhishek ^{[4
]}

Kumar, Vipul ^{[5
]}

Guliani, Astha ^{[5
]}

Kumar, Nitin ^{[6
]}

Hooda, Vikas ^{[1
]}

Yadav, Aparna ^{[7
]}

Nehra, Kiran ^{[2
]}

Mehta, Promod K. ^{[1
,3
,8
]}

机构：

[1] Maharshi Dayanand Univ MDU, Ctr Biotechnol, Rohtak, India

[2] Deenbandhu Chhotu Ram Univ Sci & Technol DCRUST, Dept Biotechnol, Sonipat, India

[3] Shree Guru Gobind Singh Tricentenary SGT Univ, Fac Allied Hlth Sci, Microbiol Dept, Gurugram, India

[4] Univ Delhi, Ramanujan Coll, Dept Stat, New Delhi, India

[5] Univ Hlth Sci UHS, Dept TB & Resp Med, Rohtak, India

[6] MDU, Ctr Med Biotechnol, Rohtak, India

[7] UHS, Microbiol Dept, Rohtak, India

[8] SGT Univ, Fac Allied Hlth Sci, Microbiol Dept, Gurugram 122505, India

来源：

EXPERT REVIEW OF RESPIRATORY MEDICINE | 2023年 / 17卷 / 11期

关键词：

Diagnosis; dry methyl green; GeneXpert; multi-targeted LAMP; Mtb; pleural TB; SYBR Green I; SYBR Green I plus dUTP-UNG assay; EXTRAPULMONARY TUBERCULOSIS; MYCOBACTERIUM-TUBERCULOSIS; LAMP; PCR; PULMONARY; IS6110;

D O I：

10.1080/17476348.2023.2292738

中图分类号：

R56 [呼吸系及胸部疾病];

学科分类号：

摘要：

Background: Diagnosis of pleural tuberculosis (TB) is tedious owing to its close resemblance with malignant pleural effusion and sparse bacterial load in clinical specimens. There is an immediate need to design a rapid and dependable diagnostic test to prevent unnecessary morbidity/mortality.Research design and methods: A multi-targeted loop-mediated isothermal amplification (MT-LAMP) was deliberated using mpt64 and IS6110 to diagnose pleural TB within pleural fluids/biopsies. MT-LAMP products were analyzed by gel-based and visual detection methods, viz. SYBR Green I, SYBR Green I+deoxyuridine triphosphate uracil-N-glycosylase (dUTP-UNG), and dry methyl green reactions.Results: In a pilot study, while assessing pleural TB/non-TB control subjects (n = 40), both SYBR Green I+dUTP-UNG/gel-based MT-LAMP assays exhibited better sensitivity/specificity than SYBR Green I and dry methyl green MT-LAMP. Since it is facile to work with SYBR Green I+dUTP-UNG than gel-based MT-LAMP, we validated the performance of SYBR Green I+dUTP-UNG in a higher number of specimens (n = 97), which revealed somewhat higher sensitivity (85.2 vs. 81.5%) and specificity (97.7 vs. 90.7%) than SYBR Green I MT-LAMP. Furthermore, the sensitivity attained by SYBR Green I+dUTP-UNG MT-LAMP was significantly higher (p < 0.001) than GeneXpert.Conclusions: Our SYBR Green I+dUTP-UNG MT-LAMP is a simple and reliable method to diagnose pleural TB, which may translate into a point-of-care test.

引用

页码：1079 / 1089

页数：11