Increased buffer pH enhances sensitivity and specificity of human papillomavirus detection using consensus primer based PCR.
Polymerase chain reaction (PCR) buffers were optimized for the specific detection of human papillomavirus (HPV) sequences. The effect of pH, potassium chloride concentration and magnesium chloride concentration of three different consensus primers were examined. Several phylogenetically distinct HPVs (HPV1, HPV2, HPV6, HPV8, HPV16, HPV18, and HPV20) were used to determine the optimal buffer components. Genital types were less sensitive to changes in pH than cutaneous types. Higher buffer pH, with a few exceptions, led to increased sensitivity and specificity of HPV detection.
Payne
D
D
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston 77555-1029, USA.
Hoskins
S
S
Schouten
H
H
van Vleuten
H
H
Tyring
S
S
D001483:Base Sequence; D002021:Buffers; D016384:Consensus Sequence; D017931:DNA Primers; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015636:Magnesium Chloride; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D011189:Potassium Chloride / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014412:Tumor Virus Infections / Q000175:diagnosis
D002021:Buffers; D017931:DNA Primers; D015636:Magnesium Chloride; D011189:Potassium Chloride