[Detection and genotyping of human papillomavirus (HPV) using PCR-LIS-SSCP].
This study describes a fast and simple method for human papillomavirus (HPV) typing based on the polymerase chain reaction (PCR) amplification of a portion of the viral genome and single strand conformation polymorphism using low ionic strength solutions (LIS-SSCP). PCR products were obtained using My09/My11 and Gp5/Gp6 primers in a nested reaction. The band patterns corresponded to the plasmid HPV clones from HPV-6, -11, -16, -18, -31, -33 and -34. The SSCP minigels were stained with SYBR-Green II. In order to determine diagnostic applicability, 100 cervical samples were studied comprising liquid cytology and paraffin embedded biopsies from patients showing squamous intraepithelial lesions (SILs). The SSCP patterns obtained from the clinical samples and the HPV clones were similar when the same type was present. Therefore, the methodology proved to be efficient and with high reproducibility for the detection and typing of HPV in clinical samples.
Gomez
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MA
Centro de Investigaciones en Genética Básica y Aplicada (CIGEBA), Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, Calle 60 y 118 s/n, 1900 La Plata, Argentina. magomez@fcv.medvet.unlp.edu.ar
Abba
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MC
Golijow
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CD
D001706:Biopsy; D002294:Carcinoma, Squamous Cell / Q000821:virology; D002584:Cervix Uteri / Q000821:virology*; D004396:Coloring Agents; D004273:DNA, Neoplasm / Q000302:isolation & purification; D004279:DNA, Viral / Q000302:isolation & purification; D005260:Female; D005456:Fluorescent Dyes; D016679:Genome, Viral; D005838:Genotype; D006801:Humans; D007038:Hypotonic Solutions; D009691:Nucleic Acid Denaturation; D009930:Organic Chemicals; D009994:Osmolar Concentration; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000821:virology*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D012680:Sensitivity and Specificity; D013048:Specimen Handling; D014412:Tumor Virus Infections / Q000821:virology*; D002583:Uterine Cervical Neoplasms / Q000821:virology; D002575:Uterine Cervicitis / Q000821:virology; D014626:Vaginal Smears; D002578:Uterine Cervical Dysplasia / Q000821:virology
D004396:Coloring Agents; D004273:DNA, Neoplasm; D004279:DNA, Viral; D005456:Fluorescent Dyes; D007038:Hypotonic Solutions; D009930:Organic Chemicals; C098798:SYBR Green II