Rapid detection of hepatitis C virus RNA by a reverse transcription loop-mediated isothermal amplification assay

Wang, Qinqin; Zhang, Jie; Hu, Jinsong; Chen, Hao-tai; Du, Li; Wu, Li-qin; Ding, Yongsheng; Xiong, Sheng-he; Huang, Xin-cheng; Zhang, Yinhong; Liu, Yongsheng

doi:10.1111/j.1574-695x.2011.00828.x

Cited by 15 publications

(20 citation statements)

References 20 publications

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“…These genotype‐specific primers provide higher specificity over conventional PCR, which only use two primers for amplification. The specificity of our RT‐LAMP was 100%, which was consistent with previous reports …”

Section: Discussionsupporting

confidence: 92%

“…The advantages of RT‐LAMP are the same as LAMP, but the template used is RNA instead of DNA . Three related RT‐LAMP assays have been reported for HCV‐RNA detection but without genotype identification . In this context, Nyan and Swinson first designed the RT‐LAMP method for identification of HCV genotypes 1a, 1b, 3, 4a, 5a, and 6a; however, their RT‐LAMP assay did not investigate genotype 2a due to limitations with the experimental materials, and it is important to note that Nyan and Swinson used GelGreen (a DNA intercalating dye) to visualize the amplified products.…”

Section: Introductionmentioning

confidence: 99%

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Detection of HCV genotypes 1b and 2a by a reverse transcription loop‐mediated isothermal amplification assay

Zhao

Liu

Sun

2017

Journal of Medical Virology

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Hepatitis C virus (HCV) genotypes 1b and 2a are the major cause of liver disease in northern China; however, conventional detection tools are labor-consuming, technically demanding, and costly. Here, we assessed the specificity, sensitivity, and clinical utility of reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of HCV genotypes 1b and 2a. Firstly, clinical samples were collected from HCV genotype 1b and 2a infected patients and the RNA were extracted. Secondly, specificity of RT-LAMP assay for detection HCV genotypes 1b and 2a were tested against viral genomes of other hepatitis viruses. Sensitivity of RT-LAMP assay was determined using serial dilutions of standard HCV genotypes 1b and 2a. The amplified products were detected by both electrophoresis and calcein/Mn -dependent visual methods. Finally, we compared the clinical detection rate of RT-LAMP to that of real-time PCR. RT-LAMP assay showed high specificity to detect HCV genotypes 1b and 2b since there was no cross-reactivity with other hepatitis viruses. Sensitivity of RT-LAMP was 100 IU/mL for both genotypes detected by either electrophoresis or calcein/Mn -dependent visual methods. The detection rate of RT-LAMP assay in clinical samples was also comparable to that of real-time PCR without significant difference between the both assays. This study proposes a newly developed RT-LAMP assay for detection of HCV genotypes 1b and 2a. RT-LAMP is highly specific, sensitive, and simple diagnostic tool which would be useful for screening and early diagnosis of HCV especially in resource-limited environments.

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Section: Discussionsupporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Detection of HCV genotypes 1b and 2a by a reverse transcription loop‐mediated isothermal amplification assay

Zhao

Liu

Sun

2017

Journal of Medical Virology

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“…103,104,106 RT-LAMP uses between four and seven primers to recognize multiple complementary regions within the 5′ UTR of the viral genome, generating high-molecular-weight concatenated amplicons. These amplicons can be detected via a visual turbidimetric readout with fluorescence enhancement.…”

Section: Amplification and Detectionmentioning

confidence: 99%

“…103,104 TMA-based HCV assays are commercialized by Hologic. TMA and NASBA use RNA polymerasemediated amplification, in contrast to PCR and LAMP.…”

Section: Amplification and Detectionmentioning

confidence: 99%

Diagnosis and Management of Hepatitis C Virus Infection

et al. 2015

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The hepatitis C virus (HCV) infects more than 200 million people globally, with increasing incidence, especially in developing countries. HCV infection frequently progresses to chronic liver disease, creating a heavy economic burden on resourcepoor countries and lowering patient quality of life. Effective HCV diagnosis, treatment selection, and treatment monitoring are important in stopping disease progression. Serological assays, which detect anti-HCV antibodies in the patient after seroconversion, are used for initial HCV diagnosis. Qualitative and quantitative molecular assays are used to confirm initial diagnosis, determine viral load, and genotype the dominant strain. Viral load and genotype information are used to guide appropriate treatment. Various other biomarker assays are performed to assess liver function and enable disease staging. Most of these diagnostic methods are mature and routinely used in high-resource countries with well-developed laboratory infrastructure. Few technologies, however, are available that address the needs of low-resource areas with high HCV prevalence, such as Africa and Southeast Asia.

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“…Since 2000, that Notomi and colleagues have developed LAMP technique, it has been widely used for diagnosis and quantification of viral pathogens. LAMP is a potent instrument for DNA amplification in detection of HBV (36, 37) which can also be used for detection of RNA hepatitis viruses (HAV, HCV and HEV) in reverse transcription way (38). Low time consumption, isothermal conditions and also no need for specific instruments are of this method's advantages (39).…”

Section: Resultsmentioning

confidence: 99%

Classical and Modern Approaches Used for Viral Hepatitis Diagnosis

2014

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Context:Viral hepatitis diagnosis is an important issue in the treatment procedure of this infection. Late diagnosis and delayed treatment of viral hepatitis infections can lead to irreversible liver damages and occurrence of liver cirrhosis and hepatocellular carcinoma. A variety of laboratory methods including old and new technologies are being applied to detect hepatitis viruses. Here we have tried to review, categorize, compare and illustrate the classical and modern approaches used for diagnosis of viral hepatitis.Evidence Acquisition:In order to achieve a comprehensive aspect in viral hepatitis detection methods, an extensive search using related keywords was done in major medical library and data were collected, categorized and summarized in different sections.Results:Analyzing of collected data resulted in the wrapping up the hepatitis virus detection methods in separate sections including 1) immunological methods such as enzyme immunoassay (EIA), radio-immunoassay (RIA) immuno-chromatographic assay (ICA), and immuno-chemiluminescence 2) molecular approaches including non-amplification and amplification based methods, and finally 3) advanced biosensors such as mass-sensitive, electrical, electrochemical and optical based biosensors and also new generation of detection methods.Conclusions:Detection procedures in the clinical laboratories possess a large diversity; each has their individual advantages and facilities' differences.

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Rapid detection of hepatitis C virus RNA by a reverse transcription loop-mediated isothermal amplification assay

Cited by 15 publications

References 20 publications

Detection of HCV genotypes 1b and 2a by a reverse transcription loop‐mediated isothermal amplification assay

Detection of HCV genotypes 1b and 2a by a reverse transcription loop‐mediated isothermal amplification assay

Diagnosis and Management of Hepatitis C Virus Infection

Classical and Modern Approaches Used for Viral Hepatitis Diagnosis

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