Comparison of the polymerase chain reaction and Southern blot analysis in detecting and typing human papilloma virus deoxyribonucleic acid in tumors of the lower female genital tract.
To conduct studies on the clinical and pathologic significance of human papilloma virus (HPV) in genital malignancies, accurate detection and typing of the virus in clinical material are essential. Currently, Southern blotting and the polymerase chain reaction (PCR) are two of the most commonly used methods to identify HPV. This study was undertaken to compare these techniques in the detection and typing of HPV in 242 invasive malignancies of the lower female genital tract. BamHI and PstI restriction digests of tumor DNA were hybridized to 32P-labeled probes for HPV types 6, 16, and 18 at TM -20 degrees C after Southern transfer. Blots were then washed at Tm -20 degrees C and Tm -9 degrees C. The DNA was also amplified by PCR using both highly conserved consensus L1 primers that detect 25 different HPV genotypes and primers specific for HPV 6 E6, 16 E7, and 18 E6. All PCR products were hybridized to type-specific radiolabeled probes. In 202 of the 242 (83%) samples, HPV was detected, including 189 of 218 (87%) cervical cancers, 11 of the 20 (55%) vulvar cancers, and two of four tumors from the vagina, urethra, or anus. In 67% of the specimens, there was agreement between the Southern blot technique and both methods of PCR (consensus and type-specific primers), including 121 of the 202 HPV-positive specimens and 40 HPV-negative specimens. Of the 141 tumors with HPV detected by Southern blot analysis, the same HPV type was detected by PCR in 121 (86%).(ABSTRACT TRUNCATED AT 250 WORDS)
Monk
B J
BJ
Department of Obstetrics and Gynecology, UCI Medical Center, University of California, Irvine, Orange.
Cook
N
N
Ahn
C
C
Vasilev
S A
SA
Berman
M L
ML
Wilczynski
S P
SP
D001483:Base Sequence; D015139:Blotting, Southern / Q000379:methods*; D002277:Carcinoma / Q000235:genetics; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D005833:Genital Neoplasms, Female / Q000235:genetics / Q000821:virology*; D006801:Humans; D008969:Molecular Sequence Data; D009856:Oncogene Proteins, Viral / Q000235:genetics; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012703:Serotyping
D017931:DNA Primers; D004279:DNA, Viral; D009856:Oncogene Proteins, Viral