Detection of human papilloma virus in clinical samples by polymerase chain reaction (PCR)

Sub-title
AuthorsM Cassar
C Vidal
AbstractAim: The Human Papilloma viruses (HPV) are a group of viruses that includes more than a hundred types, thirty of whom were reported to be transmitted sexually and infect the genital area in both males and females. Detection of HPV is very difficult since it might be present very high up in the vagina, cervix or anus. Until now observation of koilocytosis in routing Pap smear is indicative of HPV infection although this is not always the case. In this study a molecular test for the detection of HPV infection was developed. Method: DNA was extracted from cervical brushings or biopsies by conventional methods. HPV DNA controls of different types including 16, 18 and non-oncogenic types were also used in the study. A set of primers consisting of nine pairs was used for PCR amplification. In parallel, another PCR was performed as a control to test for the efficiency of the extraction method. Following PCR, the expected fragment of approximately 400bp was detected by agarose gel electrophoresis. Results: Positive amplification was observed for all HPV controls and known positive samples. DNA sequencing was then performed using BigDye terminator technique and results compared to public databases to identify the type. There was 100% concordance with the expected types of HPV controls. Conclusions: In this study a novel method for detection and typing of HPV in clinical samples was developed. This method is very sensitive and specific and can detect early infection with this virus and so help in better treatment management.

Published in:
JournalMalta Medical Journal
Volume15 Issue 1-2/suppl. 2003
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Date
Link to journal

Key wordsPCR, human papilloma virus

Compiled by: Dr. I. Stabile    Dr. J. Pace