[Show abstract][Hide abstract] ABSTRACT: A peptide nucleic acid (PNA) probe-based real-time PCR (PNAqPCR™ TB/NTM detection kit; PANAGENE, Korea) assay has been recently developed for the simultaneous detection of Mycobacterium tuberculosis complex (MTBC) and nontuberculous mycobacteria (NTM) in clinical specimens. The study was aimed at evaluation of the performance of PNA probe-based real-time PCR in respiratory specimens.
To evaluate potential cross-reactivity, the extracted DNA specimens from Mycobacterium species and non-mycobacterial species were tested using PNA probe-based real-time PCR assay. A total of 531 respiratory specimens (482 sputum specimens and 49 bronchoalveolar washing fluid specimens) were collected from 230 patients in July and August, 2011. All specimens were analyzed for the detection of mycobacteria by direct smear examination, mycobacterial culture, and PNA probe-based real-time PCR assay.
In cross-reactivity tests, no false-positive or false-negative results were evident. When the culture method was used as the gold standard test for comparison, PNA probe-based real-time PCR assay for detection of MTBC had a sensitivity and specificity of 96.7% (58/60) and 99.6% (469/471), respectively. Assuming the combination of culture and clinical diagnosis as the standard, the sensitivity and specificity of the new real-time PCR assay for detection of MTBC were 90.6% (58/64) and 99.6% (465/467), respectively. The new real-time PCR for the detection of NTM had a sensitivity and specificity of 69.0% (29/42) and 100% (489/489), respectively.
The new real-time PCR assay may be useful for the detection of MTBC in respiratory specimens and for discrimination of NTM from MTBC.
Annals of Laboratory Medicine 07/2012; 32(4):257-63. · 1.48 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A major problem of long-term antiviral therapy in chronic hepatitis B patients is the emergence of hepatitis B virus (HBV) mutations associated with drug resistance. Recently, a new array using peptide nucleic acids (PNAs), which are synthetic nucleic acid analogues, was developed for the detection of HBV mutations at six different codon positions associated with lamivudine (LAM) and adefovir (ADV) resistance. We compared the PNA array with direct sequencing and reverse hybridization (INNO-LiPA) in 73 samples obtained from chronic hepatitis B patients. The PNA array detected mutations associated with LAM and/or ADV resistance in 60 (82.2%) of the 73 samples. The overall concordance rate of PNA array and INNO-LiPA compared with direct sequencing was 99.5% and 98.2%, respectively. The rate of complete concordance between PNA array and INNO-LiPA was 92.7%. The PNA array assay results were comparable with INNO-LiPA for detection of HBV mutations associated with antiviral resistance.
Archives of Virology 05/2011; 156(9):1517-24. · 2.28 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Accurate and rapid diagnosis of Pandemic Influenza A/H1N1 2009 virus (H1N1 2009) infection is important for the prevention and control of influenza epidemics and the timely initiation of antiviral treatment. This study was conducted to evaluate the performance of several diagnostic tools for the detection of H1N1 2009. Flocked nasopharyngeal swabs were collected from 254 outpatients of suspected H1N1 2009 during October 2009. This study analyzed the performances of RealTime ready Inf A/H1N1 Detection Set (Roche), Influenza A (H1N1) Real-Time Detection Kit (Bionote), Seeplex Influenza A/B OneStep Typing set (Seeplex reverse transcriptase PCR [RT-PCR]), BinaxNow Influenza A & B test kit (Binax rapid antigen test [RAT]) and SD BIOLINE Influenza Ag kit (SD RAT). Roche and Bionote real-time RT-PCR showed identical results for the H1N1 2009 hemagglutinin gene. Compared with real-time RT-PCR, the sensitivities and specificities were 83.7% and 100% for Seeplex RT-PCR, 64.5% and 94.7% for Binax RAT, and 69.5% and 100% for SD RAT. The sensitivities of Seeplex RT-PCR, Binax RAT and SD RAT in patients aged over 21 years were 73.7%, 47.4% and 57.9%, respectively. The sensitivities of Seeplex RT-PCR, Binax RAT and SD RAT on the day of initial symptoms were mostly lower (68.8%, 56.3% and 31.3%, respectively). In conclusion, multiplex RT-PCR and RAT for the detection of H1N1 2009 were significantly less sensitive than real-time RT-PCR. Also, a negative RAT may require more sensitive confirmatory assays, because it cannot be ruled out from influenza infection.
Journal of Microbiology and Biotechnology 10/2010; 20(10):1450-6. · 1.40 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We evaluated the SD Bioline Influenza Ag A/B/A(H1N1) Pandemic test kit and compared it with real-time reverse transcriptase PCR (RT-PCR) for its ability to detect H1N1 2009. The sensitivity and specificity of the test kit for H1N1 2009 were 77% and 100%, respectively.
Journal of clinical microbiology 03/2010; 48(6):2260-2. · 4.16 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We compared the sensitivities and specificities of peptide nucleic acid (PNA) probe-based real-time PCR (PNAqPCR™ TB/NTM) and Cobas TaqMan MTB assays for detection of Mycobacterium tuberculosis complex (MTBC) in clinical specimens. A total of 425 clinical specimens including 360 respiratory specimens and 65 non-respiratory specimens were evaluated for comparative analysis. In respiratory specimens, the sensitivity of TaqMan MTB and PNAqPCR assay for detection of MTBC was 82.9% and 91.5%, respectively. In non-respiratory specimens, the sensitivity of the TaqMan MTB and PNAqPCR assay was 23.1% and 76.9%, respectively. Overall, the sensitivity and specificity of the TaqMan MTB assay for detection of MTBC was 76.9% and 100%, respectively. The PNAqPCR assay had a sensitivity and specificity of 90% and 99.7%, respectively.