LNA real-time PCR probe quantification of hepatitis B virus DNA

Wang, Qing; Wang, Xueqian; Zhang, Junhua; Song, Guanghui

doi:10.3892/etm.2011.442

Cited by 14 publications

(8 citation statements)

References 18 publications

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“…Padlock probes were used in the rolling circle amplifications, which required complex procedure [ 18 ]. In addition to above probes, LNA probe is another kind of probe that has been widely used in real-time PCR to quantitatively detect pathogens [ 20 , 21 ]. The LNA probe contains oligonucleotide monomers modified with an additional methylene bridge between 2′ oxygen and 4′ carbon of the ribose ring.…”

Section: Introductionmentioning

confidence: 99%

Locked Nucleic Acid Probe-Based Real-Time PCR Assay for the Rapid Detection of Rifampin-Resistant Mycobacterium tuberculosis

Zhao

Li²,

Sun

et al. 2015

PLoS ONE

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Drug-resistant Mycobacterium tuberculosis can be rapidly diagnosed through nucleic acid amplification techniques by analyzing the variations in the associated gene sequences. In the present study, a locked nucleic acid (LNA) probe-based real-time PCR assay was developed to identify the mutations in the rpoB gene associated with rifampin (RFP) resistance in M. tuberculosis. Six LNA probes with the discrimination capability of one-base mismatch were designed to monitor the 23 most frequent rpoB mutations. The target mutations were identified using the probes in a “probe dropout” manner (quantification cycle = 0); thus, the proposed technique exhibited superiority in mutation detection. The LNA probe-based real-time PCR assay was developed in a two-tube format with three LNA probes and one internal amplification control probe in each tube. The assay showed excellent specificity to M. tuberculosis with or without RFP resistance by evaluating 12 strains of common non-tuberculosis mycobacteria. The limit of detection of M. tuberculosis was 10 genomic equivalents (GE)/reaction by further introducing a nested PCR method. In a blind validation of 154 clinical mycobacterium isolates, 142/142 (100%) were correctly detected through the assay. Of these isolates, 88/88 (100%) were determined as RFP susceptible and 52/54 (96.3%) were characterized as RFP resistant. Two unrecognized RFP-resistant strains were sequenced and were found to contain mutations outside the range of the 23 mutation targets. In conclusion, this study established a sensitive, accurate, and low-cost LNA probe-based assay suitable for a four-multiplexing real-time PCR instrument. The proposed method can be used to diagnose RFP-resistant tuberculosis in clinical laboratories.

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Section: Introductionmentioning

confidence: 99%

Locked Nucleic Acid Probe-Based Real-Time PCR Assay for the Rapid Detection of Rifampin-Resistant Mycobacterium tuberculosis

Zhao

Li²,

Sun

et al. 2015

PLoS ONE

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“…The limit of the duplex real-time PCR assay was 29.5 IU/ml, whereas the specificity was 100% for the detection of HBV DNA [137]. A trial has been tested, a TaqMan locked nucleic acid (LNA) real-time polymerase chain reaction (PCR) probe for the accurate quantification and detection of hepatitis B virus (HBV) DNA in serum (plasma) [138]. The genotyping analysis of HBV can performed by realtime PCR using (GQ-PCR) method or the direct sequencing and reverse hybridization with INNO-LiPA HBV genotyping assay [139].…”

Section: Viral Hepatitismentioning

confidence: 99%

Molecular Diagnostics as an Indispensable Tool for the Diagnosis of Infectious Diseases of Viral Origin and Global Impact

Rosas-Murrieta¹,

Herrera‐Camacho²,

Peña³

et al. 2014

Trends in Infectious Diseases

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“…for the humoral immune response and is believed to play a vital role in viral entry into host cells [Solomon et al, 2003]. The xlink protein is involved in virus replication and regulation of the innate immune response [Li et al, 2012;Zhang et al, 2012]. The functions of NS3 and NS4 are prominent, they code for serine protease and RNA dependent RNA polymerase (RdRp) [Lu et al, 2013].…”

Section: Viral Replication and Morphogenesismentioning

confidence: 99%

Trends in Infectious Diseases

Saxena¹

2014

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is a Medical Microbiologist at CSIR-Centre for Cellular and Molecular Biology (CCMB) in India. The main research interests of his group are to understand the epidemiology and molecular mechanisms of host-defense during human viral infections and to develop new predictive, preventive and therapeutic strategies for them using JEV and HIV as a model. His research work has been published in various high impact factor journals with high citation. He has received many awards and honors in India and abroad including various Young Scientist Awards, BBSRC India Partnering Award and named as "Global Leader in Science" by The Scientist magazine (USA).

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LNA real-time PCR probe quantification of hepatitis B virus DNA

Cited by 14 publications

References 18 publications

Locked Nucleic Acid Probe-Based Real-Time PCR Assay for the Rapid Detection of Rifampin-Resistant Mycobacterium tuberculosis

Locked Nucleic Acid Probe-Based Real-Time PCR Assay for the Rapid Detection of Rifampin-Resistant Mycobacterium tuberculosis

Molecular Diagnostics as an Indispensable Tool for the Diagnosis of Infectious Diseases of Viral Origin and Global Impact

Trends in Infectious Diseases

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