Comparison of enzyme immunoassay, shell vial culture, and conventional cell culture for the rapid detection of herpes simplex virus

Five hundred specimens (288 genital, 192 dermal, and 20 ocular) were extracted by technologists, and the DNA was assayed by LightCycler PCR (DNA polymerase and thymidine kinase [TK] gene targets) and by conventional tube and shell vial cell culture. One hundred fifty-eight confirmed (by cell culture and TK target PCR) positive and LightCycler-positive specimens were detected during the first 30 PCR cycles. LightCycler PCR-positive results for cycles 31 to 45 (39 of 67 [58.2%]) required confirmation by another PCR target (TK). LightCycler PCR is more sensitive (n = 197; 23.1%) than cell cultures (n = 150) for the routine laboratory detection of herpes simplex virus infections.

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Capillary enzyme immunoassay for rapid detection of herpes simplex virus in clinical specimens.

Journal of Clinical Microbiology ◽

10.1128/jcm.25.2.320-322.1987 ◽

1987 ◽

Vol 25 (2) ◽

pp. 320-322 ◽

Cited By ~ 5

Author(s):

I C Shekarchi ◽

D A Fuccillo ◽

R Strouse ◽

J L Sever

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Enzyme Immunoassay ◽

Rapid Detection ◽

Clinical Specimens ◽

Simplex Virus

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Diagnosis of Herpes Simplex Virus Infections in the Clinical Laboratory by LightCycler PCR

Journal of Clinical Microbiology ◽

10.1128/jcm.38.2.795-799.2000 ◽

2000 ◽

Vol 38 (2) ◽

pp. 795-799 ◽

Cited By ~ 143

Author(s):

Mark J. Espy ◽

James R. Uhl ◽

P. Shawn Mitchell ◽

Jill N. Thorvilson ◽

Kathleen A. Svien ◽

...

Keyword(s):

Cell Culture ◽

Herpes Simplex Virus ◽

Herpes Simplex ◽

Cell Cultures ◽

Melting Curve ◽

Clinical Manifestations ◽

Melting Curve Analysis ◽

Shell Vial ◽

Pcr Products ◽

Simplex Virus

Herpes simplex virus (HSV) causes several clinical manifestations in both normal and immunocompromised hosts; this agent is the most frequently detected virus in diagnostic laboratories. Recovery of the virus in cell culture is considered the “gold standard” for detection of this virus from sources other than cerebrospinal fluid. LightCycler is a newly developed, commercially available system designed to rapidly perform PCR, with real-time detection of PCR products by a fluorescence resonance energy transfer assay. We compared the detection of HSV for 200 specimens (number of genital specimens, 160; number of dermal specimens, 38; number of ocular specimens, 2) by shell vial cell cultures (MRC-5) and by LightCycler PCR. Of a total of 88 (44%) HSV strains detected, 69 (78%) were detected by both shell vial cell cultures and LightCycler PCR (DNA polymerase target). A total of 19 (22%) specimens were detected exclusively by LightCycler PCR. No specimens were positive by the shell vial assay only. All 19 discrepant samples had HSV DNA detected by an independent PCR directed to the thymidine kinase gene of the virus. The melting curve analysis feature of the LightCycler instrument identified identical genotype results for HSV type 1 (HSV-1) and HSV-2 from 84 of 88 (96%) positive samples. Specimens can be extracted, target HSV DNA can be amplified, and HSV PCR products can be identified by genotype within 2 h after receipt of specimen into the laboratory. The increased level of accurate identification (all 88 positive samples) compared with that of shell vial cell culture (69 of 88 samples identified as positive) and the agreement of LightCycler PCR results with all shell vial positive results indicate the potential for routine implementation of this technology for laboratory diagnosis of HSV infections.

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