A simple HPV 18 detection method based on ultra specific primer immobilized on glass slides.
This study was carried out to develop a simple and inexpensive method for detection of Human papillomavirus (HPV 18) based on irreversible immobilization of ultra specific primer on silanized glass slides. This method is revealed by Blue Green Loading Dye I (LGC) and compared with conventional polymerase chain reaction (PCR) for endocervical samples. The new method was tested in 40 DNA samples with precancer uterine lesions of women treated in Hospital of Recife PE, Brazil. DNA samples were extracted using Wizard Genomic DNA Purification Kit according to manufacturer's instructions. The samples were tested for HPV 18 by conventional PCR (PCRc) and the products visualized on 1.2% agarose gel, with LGC under UV 260 nm. After that the positive and negative samples to HPV 18 were tested by immobilization method and the results visualized with LGC under UV 260 nm. Both PCRc and immobilization method showed high degree of correlation (95%), whereas comparison between PCRc and immobilization method showed good correlation (100%). PCRc is widely known for detection of HPV because of its high sensitivity and efficiency, but due to high cost it is not yet standardized for use in public health laboratories. In our study, the single-stranded DNA immobilized method on a glass slide was effective in screening for HPV revealed by Blue Green and may be an alternative method for diagnosis of HPV once it offers a fast, and easy handling.
Federal Rural University of Pernambuco, Brazil.
Amanda C
AC
Fidelis
Rosa F
RF
Dutra
Paulo Roberto E
PR
Souza
Celso Pinto
CP
de Melo
Maria M D
MM
Maia
D002584:Cervix Uteri / Q000821:virology; D015331:Cohort Studies; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D005898:Glass; D052161:Human papillomavirus 18 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000821:virology*; D011230:Precancerous Conditions / Q000175:diagnosis / Q000821:virology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis / Q000821:virology
D017931:DNA Primers; D004279:DNA, Viral