Korean J Clin Pathol.  1997 Oct;17(5):764-771.

A Study on the Detection of Herpes Simplex Virus using Nested PCR

• Choi MY
• Yoo JW
• Choi TY
• Kim YT

Abstract

BACKGROUND: Herpes simplex virus (HSV) is associated with a insignificant skin lesion, keratitis, encephalitis, congenital infection, sexually transmitted disease, or cervix cancer. There are two types of serogroup, HSV-1 and HSV-2. HSV-1 makes the lesion mainly on the above-waist area and HSV-2 makes the lesion mainly on the below-waist area. To diagnose the HSV infection, immunological or cultural methods usually have been used until now. But they are not satisfactory in terms of sensitivity, specificity, and ease of application. Recently the polymerase chain reaction (PCR) was developed. Because of the exponential nature of the amplification, this method can detect extremely small amount of DNA. We compared nested PCR with cultural method for HSV detection.
METHODS
We obtained 61 specimens from the lesions of oral mucosa, face, and genital area. Samples were inoculated into the monolayer from the African green monkey kidney cell(Vero). When the slide showed cytopathic effect(CPE), HSV infection was confirmed, After extracting DNA from 61 samples, we amplified HSV DNA using nested PCR with the primers against the gene encoding glycoprotein (gD) of HSV-1 and HSV-2.
RESULTS
We found 632 bp band after the 1st PCR round and 271 bp band after the 2nd PCR round with HSV-1 specific primers. HSV-2 revealed 428 bp band after the 1st PCR round and 231 bp band after the 2nd PCR round. Nested PCR showed analytical sensitivity at 10(-9) g of DNA in HSV-1 and 10(-10) g of DNA in HSV-2. Viral culture was positive in 36%, nested PCR detected HSV DNA sequence in 54% of samples. Nested PCR typed HSV, HSV-1 in 67%, HSV-2 in 39%, and mixed type in 6% of PCR-positive samples. All isolates from above-waist area were HSV-1. Seventy seven percent of 13 isolates from below-waist area were HSV-2 and 38% were HSV-1.
CONCLUSIONS
Nested PCR offers a rapid, simple, and sensitive test for HSV infections of skin and mucosa.

MeSH Terms

Base Sequence
Cercopithecus aethiops
DNA
Encephalitis
Glycoproteins
Herpes Simplex*
Herpesvirus 1, Human
Herpesvirus 2, Human
Keratitis
Kidney
Mouth Mucosa
Mucous Membrane
Polymerase Chain Reaction*
Sensitivity and Specificity
Sexually Transmitted Diseases
Simplexvirus*
Skin
Uterine Cervical Neoplasms
DNA
Glycoproteins