ArticleJournal of clinical microbiology2000
Single-tube balanced heminested PCR for detecting Mycobacterium tuberculosis in smear-negative samples.
Article in Journal of clinical microbiology, 2000. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers, 1 of them a synthesis that pooled it.
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Who cites it
11 citing papers in PubMed, 1 synthesis or guideline pooled it, 27 citations in OpenAlex.
- In-house nucleic acid amplification tests for the detection of Mycobacterium tuberculosis in sputum specimens: meta-analysis and meta-regression.BMC microbiology · 2005Pooled it
- A Genotyping Method for Detecting Foreign Buffalo Material in Mozzarella di Bufala Campana Cheese Using Allele-Specific- and Single-Tube Heminested-Polymerase Chain Reaction.Foods (Basel, Switzerland) · 2023Article
- Performance of the BD ProbeTec ET direct detection assay for the analysis ofJournal of Taibah University Medical Sciences · 2017Article
- Performance of nested multiplex PCR assay targeting MTP40 and IS6110 gene sequences for the diagnosis of tubercular lymphadenitis.Journal of microbiology (Seoul, Korea) · 2017Article
- Diagnosis of Mycobacterium tuberculosis using molecular biology technology.Asian Pacific journal of tropical biomedicine · 2011Article
- Analysis of sequence diversity among IS6110 sequence of Mycobacterium tuberculosis: possible implications for PCR based detection.Bioinformation · 2011Article
- Development of a real-time qPCR method for detection and enumeration of Mycobacterium spp. in surface water.Applied and environmental microbiology · 2010Article
- Peptide nucleic acid array for detection of point mutations in hepatitis B virus associated with antiviral resistance.Journal of clinical microbiology · 2010Article
- Comparative evaluation of nested PCR and conventional smear methods for the detection of Mycobacterium tuberculosis in sputum samples.Molecular diagnosis & therapy · 2010Article
- Simultaneous identification of Mycobacterium genus and Mycobacterium tuberculosis complex in clinical samples by 5'-exonuclease fluorogenic PCR.Journal of clinical microbiology · 2002Article
- Rapid detection of smear-negative Mycobacterium tuberculosis by PCR and sequencing for rifampin resistance with DNA extracted directly from slides.Journal of clinical microbiology · 2001Article
Corrections and comments
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Authors and funding
6 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In order to achieve more sensitive and specific results for the rapid diagnosis of tuberculosis, we have developed a new method, named balanced heminested PCR, which avoids the inconvenience of asymmetric amplification and has the advantages of single-tube heminested PCR. This was achieved by replacing the outer primer that participates in both rounds of amplification in the standard heminested technique by another primer containing the sequence of the inner primer attached at its 5' end. When both techniques were tested for the IS6110 target of Mycobacterium tuberculosis complex in 80 smear-negative culture-positive sputum samples and 60 control samples, the results showed 100% specificity for both techniques and sensitivities of 60 and 75% for heminested PCR and balanced heminested PCR, respectively (P = 0.02). In conclusion, the balanced heminested technique shows a higher sensitivity than that of the standard heminested, and it could be applied to any PCR by attaching the inner primer at the 5' end of the opposite outer primer. Thus, the balanced heminested technique provides a target for the inner primer in both strands, avoiding asymmetric amplification and thereby resulting in a more efficient amplification, and, in practice, a higher sensitivity without loss of specificity and with a minimum risk of cross-contamination.
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