Rapid reverse transcription-PCR detection of hepatitis C virus RNA in serum by using the TaqMan fluorogenic detection system

Morris, Timothy T.; Robertson, Betty H.; Gallagher, Margaret

doi:10.1128/jcm.34.12.2933-2936.1996

Cited by 129 publications

(47 citation statements)

References 17 publications

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“…Recently, TaqMan technology has combined the 5 nuclease activity of the Taq DNA polymerase and forster resonance energy transfer to detect and quantify amplification product in a closed tube format. Using this technology real-time PCR has been developed to detect the nucleic acid [164,165]. This is most sensitive and rapid method to detect the nucleic acid.…”

Section: Real-time Polymerase Chainmentioning

confidence: 99%

A Brief Review on Diagnosis of Foot-and-Mouth Disease of Livestock: Conventional to Molecular Tools

Longjam

Deb

Sarmah

et al. 2011

Veterinary Medicine International

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Foot-and-mouth disease (FMD) is one of the highly contagious diseases of domestic animals. Effective control of this disease needs sensitive, specific, and quick diagnostic tools at each tier of control strategy. In this paper we have outlined various diagnostic approaches from old to new generation in a nutshell. Presently FMD diagnosis is being carried out using techniques such as Virus Isolation (VI), Sandwich-ELISA (S-ELISA), Liquid-Phase Blocking ELISA (LPBE), Multiplex-PCR (m-PCR), and indirect ELISA (DIVA), and real time-PCR can be used for detection of antibody against nonstructural proteins. Nucleotide sequencing for serotyping, microarray as well as recombinant antigen-based detection, biosensor, phage display, and nucleic-acid-based diagnostic are on the way for rapid and specific detection of FMDV. Various pen side tests, namely, lateral flow, RT-LAMP, Immunostrip tests, and so forth. are also developed for detection of the virus in field condition.

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Section: Real-time Polymerase Chainmentioning

confidence: 99%

A Brief Review on Diagnosis of Foot-and-Mouth Disease of Livestock: Conventional to Molecular Tools

Longjam

Deb

Sarmah

et al. 2011

Veterinary Medicine International

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“…for RTD-PCR of HBV-DNA RTD-PCR [Livak et al, 1995;Gibson et al, 1996;Heid et al, 1996;Morris et al, 1996] was performed using one of two sets of PCR primers and probes complementary to sequences located in the HBs and HBx regions . Both sets were universally conserved among all known sequences.…”

Section: Oligonucleotide Primers and Probesmentioning

confidence: 99%

Virological significance of low‐level hepatitis B virus infection in patients with hepatitis C virus associated liver disease

Tanaka

Inoue

Hayashi

et al. 2003

Journal of Medical Virology

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The clinical and virological significance of low-level viremia by hepatitis B virus (HBV) in hepatitis C virus (HCV)-infected patients remains unclear. HBV-DNA and HCV-RNA were, therefore, quantitatively analyzed in livers and sera from co-infected patients. HBV-DNA and HCV-RNA were quantitated using real-time detection of polymerase chain reaction (RTD-PCR), based on Taq-Man chemistry, in 220 non-HCV-infected healthy volunteers and 93 HCV-infected patients without detectable HBsAg. Serum HBV-DNA was detected in 4 (1.8%) of 220 non-HCV-infected healthy volunteers and 32 (34.4%) of 93 HCV-infected patients without detectable HBsAg. HCV-infected patients displayed higher frequency of HBV infection than healthy volunteers (P < 0.0001). Hepatocellular carcinoma (HCC) was more frequent among co-infected patients than among HCV mono-infected patients (P < 0.001). However, quantities of HBV-DNA in sera from co-infected patients were very low (8-19,000 copies/ml). HBV-DNA was detected in liver tissue from co-infected patients at 2-20 copies per 100 hepatocytes, accounting for 1/1,000 to 1/10,000 of HBsAg positive patients. In livers of patients with HCC and HCV or HBV mono-infection, the viruses existed predominantly in non-cancerous tissue, with levels 10- to 1,000-fold and 1- to 100-fold higher than in cancerous tissue, respectively. In contrast, patients co-infected with HCV and HBV displayed decreased HBV levels in non-cancerous tissue, but no change in cancerous tissue. These results indicate that low-level HBV infection exists in HCV-infected patients. HCC was more common among HCV/HBV co-infected patients than among HCV mono-infected patients. HCV might initiate hepatocarcinogenesis, but does not necessarily determine progression to HCC.

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“…The real-time detection of the RT-PCR products is established by targeting speci¢c release of a £uorescent reporter molecule during the PCR reaction [28^30]. During the course of PCR, processing in the primer extension by the 5P exonuclease activity of the rTth DNA polymerase degrades the internally hybridizing probe [29,30]. The degradation of the probe leads to an increase in the level of £uorescence in the reaction mixture.…”

Section: Measurement Of Viral Rna Titers By Real-time Quantitative Rementioning

confidence: 99%

A model of the real-time correlation of viral titers with immune reactions in antibody-dependent enhancement of dengue-2 infections

Chen

Yeh

Yang

et al. 2001

FEMS Immunology &Amp; Medical Microbiology

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We simultaneously assessed dengue-2 virus (DEN-2) titers by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) and immune reactions including interleukin-4 (IL-4), interferon-Q (IFN-Q) and prostaglandin E 2 (PGE 2 ) production by human mononuclear cells (MNLs) in a model of antibody-dependent enhancement (ADE). We found that DEN-1 immune sera at 1:100 and 1:250, but not those at 1:10 or control sera, enhanced DEN-2 infections in human MNLs as assessed by the fluorogenic RT-PCR technique. The enhanced profiles of DEN-2 infections determined by the RT-PCR in 6 h were reproducible by the standard plaque-forming unit (PFU) measurement established after 7 days. The ADE-enhanced DEN-2 titers determined by the RT-PCR were 5.5^33-fold higher than those detected by PFU assay, suggesting that total virions during infections were much higher than the viable ones detected by PFU assay. MNLs in response to DEN-2 infections had higher IFN-Q and PGE 2 production. However, the enhancement of DEN-2 infections by DEN-1 immune sera in MNLs was not associated with further enhancement of IFN-Q production. In contrast, the presence of subneutralizing DEN-1 immune sera that enhanced DEN-2 infections also enhanced PGE 2 but not IL-4 production. The results of this study suggest that ADE of DEN-2 infections associated with induction of immunosuppressive mediators such as PGE 2 and IL-4 can be simultaneously assessed in a real-time fashion. ß

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Rapid reverse transcription-PCR detection of hepatitis C virus RNA in serum by using the TaqMan fluorogenic detection system

Cited by 129 publications

References 17 publications

A Brief Review on Diagnosis of Foot-and-Mouth Disease of Livestock: Conventional to Molecular Tools

A Brief Review on Diagnosis of Foot-and-Mouth Disease of Livestock: Conventional to Molecular Tools

Virological significance of low‐level hepatitis B virus infection in patients with hepatitis C virus associated liver disease

A model of the real-time correlation of viral titers with immune reactions in antibody-dependent enhancement of dengue-2 infections

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