2014
DOI: 10.1016/j.diagmicrobio.2014.08.009
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Development of a highly sensitive one-tube nested real-time PCR for detecting Mycobacterium tuberculosis

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Cited by 16 publications
(12 citation statements)
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References 21 publications
(26 reference statements)
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“…tuberculosis genomic DNA, which is less than the amount of DNA in a single bacterial cell (approximately 5 fg) [ 24 ]. In the present study, this OTNPCR-LFST was 100 times more sensitive than CPCR-LFST, which is consistent with previous reports showing that nested PCR greatly increases sensitivity compared to traditional PCR [ 13 15 ]…”
Section: Discussionsupporting
confidence: 93%
See 1 more Smart Citation
“…tuberculosis genomic DNA, which is less than the amount of DNA in a single bacterial cell (approximately 5 fg) [ 24 ]. In the present study, this OTNPCR-LFST was 100 times more sensitive than CPCR-LFST, which is consistent with previous reports showing that nested PCR greatly increases sensitivity compared to traditional PCR [ 13 15 ]…”
Section: Discussionsupporting
confidence: 93%
“…Nested PCR usually takes up to 5–6 hours to complete. Investigators have attempted to develop one-tube or single-tube nested PCR technologies, in which both outer and inner primers are contained in a closed tube, the initial PCR cycles are initialed at high annealing temperatures, and the later cycles are performed at low annealing temperatures, thus reducing carry-over contamination and maintaining high sensitivity[ 13 20 ]. However, to date, this technique has been susceptible to inhibition by complex, labor-intensive operation for gel electrophoresis detection or increased equipment cost for real-time fluorescence detection, which greatly diminishes its routine use.…”
Section: Introductionmentioning
confidence: 99%
“…In recent years, polymerase chain reaction (PCR) and other methods for amplifying DNA and RNA have been developed, which enable TB diagnosis in several hours 5 7 . For example, Xpert MTB/RIF (Cepheid, Sunnyvale, CA, USA), a rapid automatic system based on real-time PCR and molecular beacon technology, used to detect M. tb genes from untreated sputum samples in less than 2 h 8 , 9 .The real-time fluorescence quantitative PCR (qPCR) was used to quantify BCG DNA in skin biopsy specimens of the BCG-vaccinated adults 10 .…”
Section: Introductionmentioning
confidence: 99%
“…The nucleic acid amplification test IS6110 has shown high levels of sensitivity to detect the presence of Mtb . One-tube nested RT-PCR is 100 times more sensitive in comparison to conventional RT-PCR ( Choi et al, 2014 ). In another study, the culture and mpt64RT-PCR demonstrated the same sensitivity (90.3%) in sputum samples.…”
Section: Current Status Of Known Biomarkers For Diagnostic Assaysmentioning
confidence: 99%