Detection and subtyping of Herpes simplex virus in clinical samples by LightCycler PCR, enzyme immunoassay and cell culture

Institute of Medical & Veterinary Science, Adelaide, South Australia.
BMC Microbiology (Impact Factor: 2.73). 07/2002; 2(1):12. DOI: 10.1186/1471-2180-2-12
Source: PubMed


Prompt laboratory diagnosis of Herpes simplex virus (HSV) infection facilitates patient management and possible initiation of antiviral therapy. In our laboratory, which receives various specimen types for detection of HSV, we use enzyme immunoassay (EIA) for rapid detection and culture of this virus. The culture of HSV has traditionally been accepted as the diagnostic 'gold standard'. In this study, we compared the use of real time PCR (LightCycler) for amplification, detection and subtyping of specific DNA with our in-house developed rapid and culture tests for HSV.
The LightCycler PCR (LC-PCR) detected and subtyped HSV in 99% (66/67) of HSV positive specimens, compared to 81% (54/67) by rapid antigen EIA or 57% (36/63) by culture. A specimen was considered positive when two or more tests yielded HSV identifications or was culture positive. Discordant results were confirmed with an in-house developed PCR-ELISA or DNA sequence analysis. The typing results obtained with the LC-PCR and by culture amplified test were completely concordant.
This study showed that the LC-PCR provided a highly sensitive test for simultaneous detection and subtyping of HSV in a single reaction tube. In addition to increased sensitivity, the LightCycler PCR provided reduced turn-around-times (2 hours) when compared to enzyme immunoassay (4 hours) or culture (4 days).

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    • "Other similarities and differences between the genomes of HSV-1 and HSV-2 are used also for genera- or type-specific molecular assays, including genes coding for some envelope glycoproteins or the DNA polymerase. The conserved gene coding for DNA polymerase is often used for the detection or quantitation of both types and based on a few mismatches between HSV-1 and HSV-2 sequences which may also be used for typing [4,5]. "
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    Virology Journal 05/2014; 11(1):83. DOI:10.1186/1743-422X-11-83 · 2.18 Impact Factor
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    • "In addition to results presented here, PCR has been used to type HSV samples on other occasions. In one study, 75 HSV-positive isolates yielded two which were untypable using type-specific antibody tests, later confirmed HSV-1 by PCR [12]. Another study yielded 1 untypable isolate of 37 tested HSV-positive isolates, which was also confirmed as HSV-1 by PCR [13]. "
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    • "Moreover, respiratory secretions were examined by real time PCR using primers and hybridization probes derived from the DNA polymerase gene of HSV [24]. Vero cell monolayers were used to isolate HSV by cell culture. "
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