Diagnosis of pulmonary and extrapulmonary tuberculosis based on detection of mycobacterial antigen 85B by immuno-PCR

被引：31

作者：

Singh, Netrapal ^{[1
]}

Sreenivas, Vishnubhatla ^{[2
]}

Gupta, Krishna B. ^{[3
]}

Chaudhary, Anil ^{[4
]}

Mittal, Anshu ^{[5
]}

Varma-Basil, Mandira ^{[5
]}

Prasad, Rajendra ^{[5
]}

Gakhar, Surender K. ^{[1
]}

Khuller, Gopal K. ^{[6
]}

Mehta, Promod K. ^{[1
]}

机构：

[1] Maharshi Dayanand Univ, Ctr Biotechnol, Rohtak 124001, Haryana, India

[2] All India Inst Med Sci, Dept Biostat, New Delhi 110029, India

[3] Univ Hlth Sci, Dept TB & Resp Med, Rohtak 124001, Haryana, India

[4] RBIPMT, Delhi 110009, India

[5] Univ Delhi, Vallabhbhai Patel Chest Inst, Delhi 110007, India

[6] Postgrad Inst Med Educ & Res, Dept Biochem, Chandigarh 160012, India

来源：

DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE | 2015年 / 83卷 / 04期

关键词：

Mycobacterium tuberculosis; Tuberculosis; SMCC; Immuno-PCR; Antigen; 85B; Diagnosis; AMPLIFIED IMMUNOASSAY; RD2; ANTIGENS; ASSAY; ANTIBODIES; COMPLEX; UTILITY; SPUTUM;

D O I：

10.1016/j.diagmicrobio.2015.08.015

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

We developed a novel indirect sandwich immuno-polymerase chain reaction (I-PCR) assay for the detection of mycobacterial antigen 85B (Ag85B, 30 kDa, Rv1886c) in pulmonary tuberculosis (PTB) and extrapulmonary tuberculosis (EPTB) patients. The amino-modified reporter DNA was covalently attached with the antidetection antibody through a heterobifunctional cross-linking agent succinimidyl 4-[N-maleimidomethyl]-cyclohexane-1-carboxylate. The detection limit of Ag85B by I-PCR was found to be 1 femtogram (fg)/mL, which was 10(6)-fold lower than an analogous enzyme-linked immunosorbent assay (ELISA). The sensitivities of 85% and 77% with I-PCR and 77.6% and 62.5% with ELISA were observed in smear-positive and smear-negative PTB patients, respectively, with high specificity. On the other hand, sensitivities of 84% and 63.7% with I-PCR and 68% and 47.5% with ELISA were observed in confirmed and clinically suspected EPTB cases, respectively, with high specificity. (C) 2015 Elsevier Inc. All rights reserved.

引用

页码：359 / 364

页数：6

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