Quantitative real-time PCR for detection of monkey B virus (Cercopithecine herpesvirus 1) in clinical samples

Cercopithecine herpesvirus 1 (monkey B virus; BV) produces extremely severe and usually fatal infections when transmitted from macaque monkeys to humans. Cercopithecine herpesvirus 16 (herpesvirus papio 2; HVP2) is very closely related to BV, yet cases of human HVP2 infection are unknown. However, following intramuscular inoculation of mice, HVP2 rapidly invades the peripheral nervous system and ascends the central nervous system (CNS) resulting in death, very much like human BV infections. In this study, the neurovirulence of HVP2 in mice was further evaluated as a potential model system for human BV infections. HVP2 was consistently neurovirulent when administered by epidermal scarification, intracranial inoculation and an eye splash. Quantitative real-time PCR, histopathology and immunohistochemistry were used to follow the temporal spread of virus following skin scarification and to compare the pathogenesis of neurovirulent and apathogenic isolates of HVP2. Apathogenic isolates were found to be capable of reaching the CNS but were extremely inefficient at replicating within the CNS. It is concluded that neurovirulent strains of HVP2 exhibit a pathogenesis in mice that parallels that observed in human BV infections and that this model system may prove useful in dissecting the viral determinants underlying the extreme severity of zoonotic BV infections.

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Sequence and genetic arrangement of the U L region of the monkey B virus ( Cercopithecine herpesvirus 1) genome and comparison with the U L region of other primate herpesviruses

Archives of Virology ◽

10.1007/s00705-003-0011-2 ◽

2003 ◽

Vol 148 (5) ◽

pp. 989-997 ◽

Cited By ~ 11

Author(s):

K. Ohsawa ◽

D. H. Black ◽

H. Sato ◽

K. Rogers ◽

R. Eberle

Keyword(s):

B Virus ◽

Herpesvirus 1 ◽

Cercopithecine Herpesvirus ◽

Monkey B Virus

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Sequence and Genetic Arrangement of the US Region of the Monkey B Virus (Cercopithecine Herpesvirus 1) Genome and Comparison with the US Regions of Other Primate Herpesviruses

Journal of Virology ◽

10.1128/jvi.76.3.1516-1520.2002 ◽

2002 ◽

Vol 76 (3) ◽

pp. 1516-1520 ◽

Cited By ~ 17

Author(s):

Kazutaka Ohsawa ◽

Darla H. Black ◽

Hiroshi Sato ◽

R. Eberle

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

B Virus ◽

The Us ◽

Herpesvirus 1 ◽

Cercopithecine Herpesvirus ◽

Simplex Virus ◽

Monkey B Virus ◽

Hsv 1

ABSTRACT The sequence of the unique short (US) region of monkey B virus (BV) was determined. The 13 genes identified are arranged in the same order and orientation as in herpes simplex virus (HSV). These results demonstrate that the BV US region is entirely colinear with that of HSV type 1 (HSV-1), HSV-2, and simian agent 8 virus.

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1176 A sensitive, quantitative real-time PCR assay to detect lamivudine resistance-associated mutations in hepatitis B virus (HBV)

Hepatology ◽

10.1016/s0270-9139(03)81214-4 ◽

2003 ◽

Vol 38 ◽

pp. 722-722

Author(s):

S LEWIN ◽

F WHITEMAN ◽

S BOWDEN ◽

A AYRES ◽

T WALTERS ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Real Time ◽

Real Time Pcr ◽

Quantitative Real Time Pcr ◽

Pcr Assay ◽

Lamivudine Resistance ◽

B Virus

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Detection of Equine Herpesvirus-1 in Nasal Swabs of Horses by Quantitative Real-Time PCR

Journal of Veterinary Internal Medicine ◽

10.1111/j.1939-1676.2008.0172.x ◽

2008 ◽

Vol 22 (5) ◽

pp. 1234-1238 ◽

Cited By ~ 24

Author(s):

G.A. Perkins ◽

L.B. Goodman ◽

E.J. Dubovi ◽

S.G. Kim ◽

N. Osterrieder

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Equine Herpesvirus ◽

Quantitative Real Time Pcr ◽

Equine Herpesvirus 1 ◽

Nasal Swabs ◽

Herpesvirus 1

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Quantitative Real-Time PCR Method for Detection of B-Lymphocyte Monoclonality by Comparison of κ and λ Immunoglobulin Light Chain Expression

Clinical Chemistry ◽

10.1373/49.1.51 ◽

2003 ◽

Vol 49 (1) ◽

pp. 51-59 ◽

Cited By ~ 88

Author(s):

Anders Ståhlberg ◽

Pierre Åman ◽

Börje Ridell ◽

Petter Mostad ◽

Mikael Kubista

Keyword(s):

B Cell ◽

Real Time ◽

Real Time Pcr ◽

B Lymphocyte ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Clinical Samples ◽

Quantitative Real Time Pcr ◽

Fine Needle ◽

Fine Needle Aspirates

Abstract Background: An abnormal IgLκ:IgLλ ratio has long been used as a clinical criterion for non-Hodgkin B-cell lymphomas. As a first step toward a quantitative real-time PCR-based multimarker diagnostic analysis of lymphomas, we have developed a method for determination of IgLκ:IgLλ ratio in clinical samples. Methods: Light-up probe-based real-time PCR was used to quantify IgLκ and IgLλ cDNA from 32 clinical samples. The samples were also investigated by routine immunohistochemical analysis and flow cytometry. Results: Of 32 suspected non-Hodgkin lymphoma samples analyzed, 28 were correctly assigned from real-time PCR measurements assuming invariant PCR efficiencies in the biological samples. Four samples were false negatives. One was a T-cell lymphoma, one was a diffuse large B-cell lymphoma, and one was reanalyzed and found lymphoma-positive by in situ calibration, which takes into account sample-specific PCR inhibition. Twelve of the samples were fine-needle aspirates, and these were all correctly assigned. Conclusions: This work is a first step toward analyzing clinical samples by quantitative light-up probe-based real-time PCR. Quantitative real-time PCR appears suitable for high-throughput testing of cancers by measuring expression of tumor markers in fine-needle aspirates.

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Establishment and Application of a Taqman Reverse Transcriptase Quantitative Real Time Pcr Assay for Feline Calicivirus

10.21203/rs.3.rs-267291/v1 ◽

2021 ◽

Author(s):

jingjie zhao ◽

Lin Liang ◽

Guangzhi Zhang ◽

Wenhui Li ◽

Shaohan Li ◽

...

Keyword(s):

Reverse Transcriptase ◽

Real Time ◽

Real Time Pcr ◽

Limit Of Detection ◽

Clinical Samples ◽

Feline Calicivirus ◽

Quantitative Real Time Pcr ◽

Pcr Assay ◽

Virus Content ◽

Nasal Swabs

Abstract Feline calicivirus (FCV) is an infectious pathogen that causes disease in cats. With the current emergence of FCV-associated virulent systemic disease (FCV VSD) worldwide, the establishment of a rapid, sensitive, and reproducible diagnostic assay for its detection is important to inform prevention and control strategies. In this study, specific primers and TaqMan-FAM probes were designed based on the conserved regions of the FCV genome sequence, and a TaqMan reverse transcriptase quantitative real time PCR assay was established. This assay could specifically detected the FCV genome. The assay had a wide dynamic range, with linear detection in the range of 9.6×109 copies/μL to 9.6×100 copies/μL, with a limit of detection of 9.6×100 copies/μL, showing high sensitivity and repeatability. In addition, we used this assay to evaluated clinical samples (n=100) taken from cats from across China for the presence/absence of FCV genetic material For samples with low virus content, the positive detection rate of TaqMan reverse transcriptase quantitative real time PCR assay (RT-qPCR) was much higher than that of conventional reverse transcriptase PCR assay (cRT-PCR). And The qRT-PCR assay was used to detect the viral load of cat swabs within 17 days after FCV infection. From days 1-9, the oral and nasal swabs generally had higher viral loads than the anal swabs. While from days 10-17, the levels in the oral and nasal swabs being generally lower than those in the anal swabs. Overall, this FCV TaqMan RT-qPCR assay assay represents a rapid and accurate.

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