Innovative Technology

Innovation

Focusing on the research and development and application of isothermal nucleic acid amplification technology

RNA恒温扩增实时检测技术与荧光定量PCR联合检测肺泡灌洗液对痰涂阴性肺结核的快速诊断价值-王静

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Abstract: Objective: To evaluate the diagnostic value of combining two distinct RNA isothermal amplification real-time PCR (FQ-PCR) methods for detecting nucleic acids from bronchoalveolar lavage fluid (BALF) specimens, particularly in patients with sputum-smear-negative pulmonary tuberculosis. Methods: A total of 1,155 BALF samples were collected from patients admitted to our hospital between November 2014 and May 2016 who were suspected of having pulmonary tuberculosis but had consistently negative sputum smears on at least three occasions. After excluding 295 cases with unclear diagnoses, 672 confirmed tuberculosis patients were assigned to the tuberculosis group, while 188 individuals with other lung diseases—including 43 cases of non-tuberculous mycobacterial infections—were included in the control group. Clinical diagnosis was used as the gold standard to calculate various evaluation metrics for SAT-TB combined with FQ-PCR testing, and these results were compared with those obtained from SAT-TB alone, FQ-PCR alone, rapid mycobacterial culture using Bactec MGIT 960, and acid-fast bacilli staining. Receiver operating characteristic (ROC) curves were plotted for comparative analysis. Results: When clinical diagnosis served as the "gold standard," ROC curve areas under the curve (AUCs) were 0.507, 0.501, 0.670, 0.660, 0.734, and 0.658 for AFS, Bactec MGIT 960, SAT-TB, FQ-PCR, parallel testing (single positive), and serial testing (double positive), respectively. Corresponding accuracies were 29.45%, 33.64%, 58.67%, 58.32%, 67.40%, and 49.82%. Notably, both SAT-TB and FQ-PCR, either individually or in combination, demonstrated statistically significant diagnostic performance (P < 0.001). Among these tests, the parallel assay showed superior diagnostic accuracy compared to SAT-TB, FQ-PCR, and serial testing; it was followed by SAT-TB, FQ-PCR, and then Bactec MGIT 960. Conclusion: Compared to conventional bacteriological methods, the combined SAT-TB and FQ-PCR approach offers several advantages for diagnosing sputum-smear-negative pulmonary tuberculosis, including rapidity, high sensitivity, and the ability to differentiate between Mycobacterium tuberculosis (MTB) and non-tuberculous mycobacteria (NTM). This method not only shortens the time required for definitive diagnosis but also enhances accuracy, making it a valuable adjunct tool for managing patients with sputum-smear-negative tuberculosis. Its clinical application is therefore highly recommended.

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探针导向重组酶介导等温扩增法检测TB_rpoB基因突变的应用价值

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重组酶介导扩增方法快速检测黄热病毒-浙江国旅

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探针导向重组酶介导等温扩增法检测TB_rpoB基因突变的应用价值