ArticleScientific reports2019
Development of a high sensitivity TaqMan-based PCR assay for the specific detection of Mycobacterium tuberculosis complex in both pulmonary and extrapulmonary specimens.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 38 citations in OpenAlex.
- Development and evaluation of a multiplex PCR for simultaneous detection of Mycobacterium tuberculosis complex, Mycobacterium avium complex, and Burkholderia pseudomallei in clinical specimens.PLoS neglected tropical diseases · 2026Article
- Validation of molecular detection ofMicrobiology spectrum · 2026Article
- Clinical characteristics of nine cases of adrenal tuberculosis and literature review.International urology and nephrology · 2026Review
- A new diagnostic kit using real-time polymerase chain reaction for the detection of Mycobacterium tuberculosis.AMB Express · 2026Article
- Partitioning and probe-based quantitative PCR assays for the wastewater monitoring ofJournal of water and health · 2025Article
- Development of SNP-LAMP Combined with Lateral Flow Dipstick to Detect the S531LDiagnostics (Basel, Switzerland) · 2025Article
- Evaluation and Comparison of Laboratory Methods in Diagnosing Mycobacterium tuberculosis and Nontuberculous Mycobacteria in 3012 Sputum Samples.The clinical respiratory journal · 2025Article
- Explore the Application Value of Metagenomic Next-Generation Sequencing of Bronchoalveolar Lavage Fluid in the Early Diagnosis of Pulmonary Tuberculosis.Infection and drug resistance · 2025Article
- Droplet digital PCR as alternative to microbiological culture forFrontiers in cellular and infection microbiology · 2024Article
- New Insights into Biomarkers for Evaluating Therapy Efficacy in Pulmonary Tuberculosis: A Narrative Review.Infectious diseases and therapy · 2023Review
- Optimization of real-time PCR protocols from lymph node bovine tissue for direct detection ofMicrobiology spectrum · 2023Article
- Facilitation of Dye-Based Quantitative Real-Time Polymerase Chain Reaction with Poly(ethylene glycol)-Engrafted Graphene Oxide.Nanomaterials (Basel, Switzerland) · 2023Article
- Review
- A Comparative Study on Visual Detection ofDiagnostics (Basel, Switzerland) · 2023Article
- Mycobacterium Tuberculosis and Avium Complex Investigation among Malaysian Free-Ranging Wild Boar and Wild Macaques at Wildlife-Livestock-Human Interface.Animals : an open access journal from MDPI · 2021Article
- Bacteria Associated with Healthcare-Associated Infections on Environmental Samples Obtained from Two Fire Departments.International journal of environmental research and public health · 2021Article
- Development of Hydrolysis Probe-Based qPCR Assays forFoods (Basel, Switzerland) · 2021Article
- Establishment of TaqMan-based real-time PCR assay for rapid detection of duck circovirus.3 Biotech · 2021Article
- Comparing mRNA expression and protein abundance in MDRBiotechnology reports (Amsterdam, Netherlands) · 2021Article
- Development of a two-step nucleic acid amplification test for accurate diagnosis of the Mycobacterium tuberculosis complex.Scientific reports · 2021Article
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Authors and funding
7 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Tuberculosis (TB) causes a heavy health burden worldwide, especially in developing countries. The need for the rapid and accurate diagnosis of TB has not been satisfied, especially for extra-pulmonary specimens or specimens with acid fast stain (AFS)-negative condition. Development and validation of a novel, sensitive and specific assay for diagnosing TB is essential. We developed IS4 primer/probe based on insertion sequence 6110 (IS6110). A qPCR assay was designed for detecting a specific region in IS6110 by BLAST. The IS4 primer/probe concentration, qPCR efficiency and various of PCR additives were evaluated and optimized. Thirty-four species of commonly isolated microorganisms were used for evaluating the analytical specificity. Moreover, 130 clinical specimens were collected for evaluating the performance versus Cobas TaqMan MTB (CTM) assay kit and culture. The amplification efficiencies of IS4 were 99.61% and 102.61% without and with internal control DNA (Bacteriophage Lambda), respectively. Dimethyl sulfoxide outperformed glycerol or BSA for eliciting the most effective amplification and the lowest limit of detection. In evaluating the clinical performance, various specimen types were collected. IS4 demonstrated a high degree of agreement (kappa = 0.71) with CTM. The clinical sensitivity and specificity of IS4 and CTM were 92.11% (35/38), 82.61% (76/92), 84.21% (32/38) and 95.65% (88/92), respectively. The clinical sensitivity and specificity of IS4 were similar for both pulmonary [92.00% (23/25) and 76.92% (30/39), respectively] and extrapulmonary [92.31% (12/13) and 86.79% (46/53), respectively] specimens. Among AFS-negative cases, the clinical sensitivity and specificity remained 90.48% (19/21) and 83.91% (73/87), respectively, with culture as the gold standard. We concluded that IS4, a new primer/probe pair for TaqMan based qPCR assay, was developed, optimized, and validated for the sensitive and specific detection of TB among various specimen types. The performance was not compromised under AFS-negative conditions.
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