Detection of herpes simplex virus DNA in cerebrospinal fluid samples using the polymerase chain reaction and microplate hybridization

Vesanen, Mika; Piiparinen, Heli; Kallio, Arja; Vaheri, Antti

doi:10.1016/0166-0934(95)01991-x

Cited by 40 publications

(30 citation statements)

References 18 publications

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“…Thirty five cycles were done in the PCR, using parameters described previously [Pitkaranta et al, 1998]. Following removal of unincorporated primers and deoxynucleoside triphosphates, hybridization of probe labeled with digoxigenin was carried out using a microplate hybridization protocol [Vesanen et al, 1996].…”

Section: Rt-pcr For Picornavirusmentioning

confidence: 99%

Picornavirus infections in children diagnosed by RT-PCR during longitudinal surveillance with weekly sampling: Association with symptomatic illness and effect of season

2006

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RT-PCR is more sensitive for rhinovirus detection than cell culture, but healthy controls are frequently rhinovirus (or picornavirus) positive in cross-sectional studies. Fifteen healthy children were followed over at least three seasons of the year with weekly sampling of nasal/nasopharyngeal secretion for RT-PCR testing for picornavirus and daily recording of respiratory symptoms. One sample positive for picornavirus was diagnosed as an infection; consecutive positive weekly samples constituted a single infection. Picornavirus illness was diagnosed if RNA was detected 7 days prior through 21 days after onset. One hundred fifty-five (21%) of 740 weekly samples were picornavirus positive and associated with illness; 37(5%) positives were not associated with illness (P = 0.001). The 192 positive samples occurred in 121 infections, 74 with a single positive and 47 with "runs" of positives in two or more consecutive samples. Forty five (96%) of the 47 runs comprised 2 or 3 consecutive positives. Ninety six (52%) of 185 reported illnesses during 235 child-months were picornavirus positive (0.4/child-month); 25 infections were asymptomatic (0.11/child-month). The infection rate was highest in fall (0.66/child-month); the winter rate (0.44/child-month) was similar to that in spring (0.5) and summer (0.43). Picornavirus infections in healthy children were common (0.51/child-month), episodic, and usually associated with brief illness; one fifth of infections were asymptomatic. The infection rate was highest in fall; infections in winter occurred at the same rate as in spring and summer.

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Section: Rt-pcr For Picornavirusmentioning

confidence: 99%

Picornavirus infections in children diagnosed by RT-PCR during longitudinal surveillance with weekly sampling: Association with symptomatic illness and effect of season

2006

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“…The amplicon was detected by hybridization in streptavidin-coated microtitre wells (Labsystems, Helsinki, Finland). Hybridization was carried out as described previously [Vesanen et al, 1996;Koskiniemi et al, 1997]. Brie¯y, 10 ml of biotinylated PCR fragment was allowed to bind onto streptavidin-coated microtitre wells.…”

Section: Real-time Pcrmentioning

confidence: 99%

Real‐time PCR for detection and quantitation of hepatitis B virus DNA

Chen

Piiparinen

Seppänen

et al. 2001

Journal of Medical Virology

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A sensitive and reproducible real-time PCR assay based on TaqMan technology was developed for the detection and quantitation of hepatitis B virus (HBV) DNA in serum, and compared with an "in-house" qualitative PCR assay. HBV DNA was measured in 125 serum samples from 76 hepatitis B patients, consisting of 22 patients with an acute infection, 20 patients with a previous history of hepatitis B infection, and 34 patients with a chronic hepatitis B. Four patients with a chronic infection were treated with either an IFN-alpha monotherapy or a combination of IFN-alpha and lamivudine. Twenty-nine sera from healthy individuals and non-hepatitis B patients served as negative controls. The assay was validated by using a 10-fold dilution series of the World Virological Quality Control (VQC) sample containing 3.73 x 10(7) genome equivalents per ml. The detection limit for the real-time PCR was 3.73 x 10(2) genome equivalents per ml (geq/ml), while it was 3.73 x 10(3) geq/ml for the in-house PCR. The real-time PCR assay had an 8-logarithm dynamic range spanning from 10(2) to 10(10) geq/ml. In clinical serum samples, the real-time PCR and the in-house PCR detected HBV DNA in 81% (101/125) and 66% (83/125) of samples, respectively. HBV DNA was not detected among the negative controls by either of these assays. In conclusion, real-time PCR is a sensitive, specific, and a reproducible approach for the detection and quantitation of HBV DNA in clinical serum samples, useful also for monitoring the efficacy of antiviral treatment.

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“…Immunoperoxidase staining for VZV was negative. The polymerase chain reaction (PCR) assay for VZV in brain tissue 9,19 gave negative results.…”

Section: Neuropathologic Findingsmentioning

confidence: 99%

Congenital Varicella-Zoster Virus Infection After Maternal Subclinical Infection: Clinical and Neuropathological Findings

et al. 2001

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Detection of herpes simplex virus DNA in cerebrospinal fluid samples using the polymerase chain reaction and microplate hybridization

Cited by 40 publications

References 18 publications

Picornavirus infections in children diagnosed by RT-PCR during longitudinal surveillance with weekly sampling: Association with symptomatic illness and effect of season

Picornavirus infections in children diagnosed by RT-PCR during longitudinal surveillance with weekly sampling: Association with symptomatic illness and effect of season

Real‐time PCR for detection and quantitation of hepatitis B virus DNA

Congenital Varicella-Zoster Virus Infection After Maternal Subclinical Infection: Clinical and Neuropathological Findings

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