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Focusing on the research and development and application of isothermal nucleic acid amplification technology
The Combined Diagnostic Value of RNA Isothermal Amplification Real-Time Detection Technology and Fluorescent Quantitative PCR for Rapid Diagnosis of Pulmonary Alveolar Lavage Fluid in Patients with Sputum-Negative Tuberculosis—Wang Jing
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Abstract: Objective: To evaluate the diagnostic value of combining two different types of nucleic acid detection methods—RNA isothermal amplification real-time detection technology (FQ-PCR)—for the simultaneous detection of target markers in bronchoalveolar lavage fluid (BALF) specimens, with a focus on its application in diagnosing sputum-negative pulmonary tuberculosis. Methods: We collected BALF specimens from 1,155 patients admitted to our hospital between November 2014 and May 2016 who were suspected of having pulmonary tuberculosis and had at least three negative sputum smear results. After excluding 295 cases with unclear diagnoses, we included 672 confirmed tuberculosis patients in the tuberculosis group and 188 patients with other lung diseases (including 43 cases of non-tuberculous mycobacterial infections) in the control group. Using clinical diagnosis as the gold standard, we calculated various evaluation indicators for the combined SAT-TB and FQ-PCR testing, and compared them with those obtained from single SAT-TB, FQ-PCR, rapid mycobacterial culture (Bactec MGIT 960), and acid-fast bacilli staining. Receiver operating characteristic (ROC) curves were plotted for comparative analysis. Results: Using clinical diagnosis as the “gold standard,” when AFS, MIGT960, SAT-TB, FQ-PCR, and the combined SAT-TB and FQ-PCR testing were used as parallel tests (single positive) or serial tests (double positive), the areas under the ROC curves (AUCs) were 0.507, 0.501, 0.670, 0.660, 0.734, and 0.658, respectively. The corresponding accuracies were 29.45%, 33.64%, 58.67%, 58.32%, 67.40%, and 49.82%. Both single and combined SAT-TB and FQ-PCR tests showed statistically significant diagnostic value (P < 0.001). In terms of diagnostic performance for sputum-negative pulmonary tuberculosis, the order of effectiveness was: parallel test > SAT-TB > FQ-PCR > serial test > sputum smear > MIGT960. Conclusion: Compared with conventional bacteriological methods, the combined SAT-TB and FQ-PCR testing demonstrates advantages in the diagnosis of sputum-negative pulmonary tuberculosis, including rapidity, sensitivity, and the ability to differentiate Mycobacterium tuberculosis (MTB) from non-tuberculous mycobacteria (NTM). This approach shortens the time to diagnosis, improves accuracy, and provides an effective auxiliary tool for patients with sputum-negative pulmonary tuberculosis, making it worthy of clinical promotion and application.
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