title: Evaluation of a convenient enzyme immunoassay to assess the quality of genital specimens submitted for the detection of human papillomavirus DNA by consensus PCR.
abstract: BACKGROUND
  In the PGMY-line blot assay, a human beta-globin fragment is co-amplified with human papillomavirus (HPV) DNA, and both analytes are detected by hybridization with probes fixed on a strip in a linear array. The beta-globin DIG-MWP test also detects beta-globin amplicons, but in a microtiter plate-based enzyme immunoassay format. Although the PGMY-line blot assay detected 50 cells per test, the beta-globin DIG-MWP test generated a signal above the detection cut-off with five cells per test.
  
  
  OBJECTIVE
  The performance of the beta-globin DIG-MWP assay to detect beta-globin DNA was assessed.
  
  
  METHODS
  The beta-globin DIG-MWP assay was compared to a standard beta-globin PCR and to the PGMY-line blot strips on 401 genital specimens. Overall, the three beta-globin assays were compared on 325 undiluted lysates, 14 diluted lysates and DNA extracted from 62 lysate samples.
  
  
  RESULTS
  Concordance between the PGMY-line blot and the standard beta-globin assay reached 99.5% (399 of 401 results), for a kappa value of 0.95. Concordant results were also obtained between the beta-globin DIG-MWP assay and PGMY-line blot assay for 387 (96.5%) of 401 test results, for a kappa value of 0.57. Discordant results were due to the increased sensitivity of the DIG-MWP assay. Using a cut-off for positivity at 1.500 optical density (OD) units for beta-globin DIG-MWP, concordance improved to 100% (401 of 401 results, kappa at 1.00).
  
  
  CONCLUSIONS
  The beta-globin DIG-MWP assay was adequate to screen for sample adequacy for HPV analysis in genital specimens.
authors.affiliation: Département de Microbiologie et Immunologie, Université de Montréal, Que., Montreal, Canada.
authors.forename: Joséphine
authors.initials: J
authors.lastname: Aho
authors.forename: Janet R
authors.initials: JR
authors.lastname: Kornegay
authors.forename: Sylvia
authors.initials: S
authors.lastname: Healey
authors.forename: Michel
authors.initials: M
authors.lastname: Roger
authors.forename: France
authors.initials: F
authors.lastname: Dion
authors.forename: Diane
authors.initials: D
authors.lastname: Gaudreault
authors.forename: Amanda P
authors.initials: AP
authors.lastname: Shepard
authors.forename: Eduardo L
authors.initials: EL
authors.lastname: Franco
authors.forename: Francois
authors.initials: F
authors.lastname: Coutlée
mesh_terms: D002584:Cervix Uteri / Q000166:cytology* / Q000821:virology; D004247:DNA / Q000032:analysis* / Q000302:isolation & purification; D004279:DNA, Viral / Q000032:analysis*; D004847:Epithelial Cells / Q000737:chemistry; D005260:Female; D005347:Fibroblasts / Q000737:chemistry; D005914:Globins / Q000235:genetics*; D006801:Humans; D007124:Immunoenzyme Techniques; D009693:Nucleic Acid Hybridization / Q000379:methods; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000453:epidemiology / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods; D011786:Quality Control; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral; D005914:Globins; D004247:DNA