Thin-layer cytology: identification of human papillomavirus by hybridization, capture and signal amplification assay using an alcoholic cell preservative medium.
OBJECTIVE
To ensure complete reliability in the detection of hyman papillomavirus using either Specimen Transport Medium (Digene Inc., Gaithersburg, Maryland, U.S.A) or an alcoholic cell preservative.


METHODS
In order to compare both media, we adjusted the cell content of specimens and optimized the denaturation duration to avoid chemiluminescence inhibition.


RESULTS
We validated this protocol in 2 groups of 90 patients, leading to agreement of 100% in both positive and negative tests. This was confirmed using linear regression curves. We demonstrated that any change in the Digene protocol, especially the use of other preservatives, should be carefully evaluated since it may interfere with DNA denaturation and hybridization and the chemiluminescence signal. The main risk was obtaining false positive responses from nonspecific DNA hybridization and false negative ones because of chemiluminescence signal inhibition.


CONCLUSIONS
It is necessary to take into account preservative characteristics when a new protocol is under validation.
Hammou
Jean-Claude
JC
Cabinet de Patologie, Hammou/Gérard, Nice, France. jchammou@wanadoo.fr
D000438:Alcohols; D002584:Cervix Uteri / Q000473:pathology / Q000821:virology; D005260:Female; D005404:Fixatives; D052163:Human papillomavirus 6 / Q000235:genetics*; D006801:Humans; D049449:Luminescence; D008163:Luminescent Measurements / Q000379:methods / Q000592:standards; D009693:Nucleic Acid Hybridization / Q000379:methods*; D030361:Papillomavirus Infections / Q000175:diagnosis*; D011237:Predictive Value of Tests; D016707:Tissue Fixation / Q000379:methods* / Q000592:standards; D002577:Uterine Cervical Diseases / Q000473:pathology / Q000821:virology*; D014626:Vaginal Smears / Q000379:methods* / Q000592:standards
D000438:Alcohols; D005404:Fixatives