Quantitation of Latent Varicella-Zoster Virus and Herpes Simplex Virus Genomes in Human Trigeminal Ganglia

Pevenstein, Stephanie R.; Williams, Russell B.; McChesney, Daniel G.; Mont, Erik K.; Smialek, John E.; Straus, Stephen E.

doi:10.1128/jvi.73.12.10514-10518.1999

Cited by 152 publications

(37 citation statements)

References 34 publications

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“…Likewise, the study of new and emerging viruses has been ideally complemented by the use of homogeneous real-time PCR assays as tools to demonstrate and strengthen epidemiological links between unique viral sequences and the [177,234,240,253,254,266,267,280,284,292,294] Solid tissues [172,198,238,[256][257][258]271,288,295] Cerebrospinal fluid [136,190,192,194,248] Peripheral blood mononuclear cells [183] Bone marrow [120] Whole blood [179,291] Plasma [118] Serum [171,172,180,279,287] Swabs [192,259,296] Bronchoalveolar lavage [243,246] Amniotic fluid [286] Saliva and sputum [158,233] Faeces [264,285] Urine [12,…”

Section: Virusesmentioning

confidence: 99%

Real-time PCR in the microbiology laboratory

Mackay

2004

Clinical Microbiology and Infection

651

338

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Use of PCR in the field of molecular diagnostics has increased to the point where it is now accepted as the standard method for detecting nucleic acids from a number of sample and microbial types. However, conventional PCR was already an essential tool in the research laboratory. Real-time PCR has catalysed wider acceptance of PCR because it is more rapid, sensitive and reproducible, while the risk of carryover contamination is minimised. There is an increasing number of chemistries which are used to detect PCR products as they accumulate within a closed reaction vessel during real-time PCR. These include the non-specific DNA-binding fluorophores and the specific, fluorophore-labelled oligonucleotide probes, some of which will be discussed in detail. It is not only the technology that has changed with the introduction of real-time PCR. Accompanying changes have occurred in the traditional terminology of PCR, and these changes will be highlighted as they occur. Factors that have restricted the development of multiplex real-time PCR, as well as the role of real-time PCR in the quantitation and genotyping of the microbial causes of infectious disease, will also be discussed. Because the amplification hardware and the fluorogenic detection chemistries have evolved rapidly, this review aims to update the scientist on the current state of the art. Additionally, the advantages, limitations and general background of real-time PCR technology will be reviewed in the context of the microbiology laboratory.

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Section: Virusesmentioning

confidence: 99%

Real-time PCR in the microbiology laboratory

Mackay

2004

Clinical Microbiology and Infection

651

338

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“…The genes encoding HSV-1 and HSV-2 glycoprotein G were selected for HSV type-specific real-time PCR. The sequences of the primers and probes used for these experiments were described by Pevenstein et al [1999]. PCR reactions were performed using the TaqMan PCR Kit (PE Applied Biosystems, Foster City, CA) according to the manufacturer's directions.…”

Section: Real-time Pcr For Hsv-1 and Hsv-2mentioning

confidence: 99%

“…Recent studies have suggested that the detection of HSV DNA by polymerase chain reaction (PCR) increases the sensitivity of detection relative to antigenic detection or cell culture methods [Cone et al, 1991[Cone et al, , 1994Lakeman and Whitley, 1995;Kimberlin et al, 1996;Mitchell et al, 1997]. While quantitative analysis of viral DNA by real-time PCR may become a valuable tool for bedside monitoring of HSV infection and progression [Pevenstein et al, 1999;Ryncarz et al, 1999;Espy et al, 2000;Kessler et al, 2000;Aldea et al, 2002;Ndjoyi-Mbiguino et al, 2003;Asano et al, 2004], it is not yet common in hospital laboratories due to the requirement of specific expensive equipment (a thermal cycler).…”

Section: Introductionmentioning

confidence: 99%

Comparison of loop‐mediated isothermal amplification, real‐time PCR, and virus isolation for the detection of herpes simplex virus in genital lesions

Sugiyama

Yoshikawa

Ihira

et al. 2005

Journal of Medical Virology

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This study compares herpes simplex virus (HSV) type-specific loop-mediated isothermal amplification (LAMP) with virus isolation and real-time PCR. Genital tract specimens were obtained from 25 patients with genital lesions; two swab samples were collected from the vulva and cervix of each patient, for a total of 50 specimens. After culturing, 10 of 50 (20%) samples were positive for HSV-1 and 12 of 50 (24%) samples were positive for HSV-2. None of the patients excreted both HSV-1 and HSV-2 virus. An original HSV type-specific LAMP assay (30 min reaction) was compared with virus isolation and HSV type-specific real-time PCR. Viral DNA was detected by LAMP in 9 of 10 HSV-1 isolated samples and 11 of 12 HSV-2 isolated samples. No viral DNA was detected in samples without virus isolation. Thus, if virus isolation was used as the standard method, the LAMP protocol was highly sensitive and specific. In comparing LAMP to real-time PCR, viral DNA was detected by the LAMP method in 9 of 12 HSV-1 DNA positive samples and 11 of 18 HSV-2 DNA positive samples. If real-time PCR was used as the standard method, then, sensitivity of the LAMP method (in particular, for HSV-2) was low. Taking this into consideration, the LAMP reaction was extended to 60 min. This led to an increase in sensitivity, resulting in an additional one and three samples testing positive for HSV-1 LAMP and HSV-2 LAMP, respectively, compared to the original LAMP protocol. Therefore, the sensitivity of the LAMP method increased to about 80%.

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“…A quantitative system was developed recently to measure the genome copies of HSV using a real-time PCR assay . This assay allows quantitation of HSV DNA accurately and reproducibly over a wide linear range [Pevenstein et al, 1999;Ryncarz et al, 1999;Ito et al, 2000]. It seems reasonable that estimation of the viral load could be used prognostically to help predict the likely course of neonatal HSV infection.…”

Section: Introductionmentioning

confidence: 99%

Quantitation of viral load in neonatal herpes simplex virus infection and comparison between type 1 and type 2

Kimura

Ito

Futamura

et al. 2002

Journal of Medical Virology

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Neonatal herpes simplex virus (HSV) infection is a severe disease with high mortality and morbidity in spite of the development of effective anti-viral therapies. The viral load in neonatal herpes simplex virus (HSV) infection was measured retrospectively in 37 patients. HSV DNA copy numbers in serum and cerebrospinal fluid (CSF) were quantified using a real-time PCR assay. Patients with disseminated infection had a higher viral load in their sera. whereas patients with central nervous system (CNS) infection exhibited a higher viral load in the CSF. The viral load was significantly higher in the serum of patients who died later. Interestingly, patients with HSV type-2 infection exhibited more CNS involvement and neurological impairment, together with a high viral load in the CSF, than did HSV type-1 patients. These results suggest that quantitation of HSV viral load may be useful for assessing the prognosis, and may provide additional information for the management of neonatal HSV infection.

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Quantitation of Latent Varicella-Zoster Virus and Herpes Simplex Virus Genomes in Human Trigeminal Ganglia

Cited by 152 publications

References 34 publications

Real-time PCR in the microbiology laboratory

Real-time PCR in the microbiology laboratory

Comparison of loop‐mediated isothermal amplification, real‐time PCR, and virus isolation for the detection of herpes simplex virus in genital lesions

Quantitation of viral load in neonatal herpes simplex virus infection and comparison between type 1 and type 2

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