Application of a new, universal DNA labeling system in the PCR mediated diagnoses of Chlamydia trachomatis and human papillomavirus type 16 infection in cervical smears.
Non-isotopic DNA labeling procedures are essential for integration of DNA diagnostics into the clinical laboratory. A newly developed reagent was tested for use in reversed hybridisation identification of DNA fragments generated by polymerase chain reaction (PCR) amplification of Chlamydia trachomatis or human papilloma virus type 16 (HPV16) DNA isolated from cervical smears. The platinum-containing chemical compound, equipped with a biotin hapten, enables versatile 'one tube' labeling of amplified DNA. A HPV16-specific probe was immobilised on a nylon strip and reverse hybridisation with the biotin labeled DNA took place. To determine the value of this new, non-isotopic label in combination with clinical material, 98 cervical smears 54 of which contained HPV16, and 51 cervical smears 26 of which contained C. trachomatis, were analysed. The novel type of non-radioactive analysis appeared to be as sensitive as its isotopic counterpart. The DNA isolation and purification method require modification only in samples of poor quality. The labeling procedure is simple, versatile and can be included as a universal linkage system in any PCR test for the detection and identification of DNA molecules.
van Belkum
A
A
Diagnostic Center SSDZ, Delft, The Netherlands.
Linkels
E
E
Jelsma
T
T
Houthoff
H J
HJ
van den Berg
F
F
Quint
W
W
D002690:Chlamydia Infections / Q000175:diagnosis* / Q000382:microbiology; D002692:Chlamydia trachomatis / Q000235:genetics* / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005069:Evaluation Studies as Topic; D005260:Female; D006801:Humans; D027383:Papillomaviridae / Q000235:genetics* / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D014412:Tumor Virus Infections / Q000175:diagnosis* / Q000382:microbiology; D002575:Uterine Cervicitis / Q000175:diagnosis* / Q000382:microbiology; D014626:Vaginal Smears; D014773:Virology / Q000379:methods / Q000706:statistics & numerical data
D004269:DNA, Bacterial; D004279:DNA, Viral