OBJECTIVE: Seven French laboratories tested the specificity and sensitivity of the polymerase chain reaction (PCR) for the detection of HIV-1 DNA. METHODS: Following its own PCR protocols, each laboratory independently tested blind two panels of 20 coded peripheral blood mononuclear cell samples collected from HIV-1-seropositive individuals and from HIV-1-seronegative individuals at high or low risk of HIV infection. For the first panel, laboratories were free to select type and number of primers; for the second, all were required to use the two primer pairs Pol 3/4 and MMy 9/10' (Nef 1). RESULTS: False-positive and false-negative results were observed in all laboratories (concordance with serology ranged from 40 to 100%). In addition, the number of positive PCR results did not differ significantly between high- and low-risk seronegatives. The use of crude cell lysates in DNA preparation produced the same PCR results as phenol-extracted DNA. Discrepancies between laboratories indicated that factors other than primer pairs contributed strongly to laboratory variability. CONCLUSIONS: Our results emphasize the importance of both positive and negative controls in PCR and demonstrate the value of multicentre PCR quality control.
OBJECTIVE: Seven French laboratories tested the specificity and sensitivity of the polymerase chain reaction (PCR) for the detection of HIV-1 DNA. METHODS: Following its own PCR protocols, each laboratory independently tested blind two panels of 20 coded peripheral blood mononuclear cell samples collected from HIV-1-seropositive individuals and from HIV-1-seronegative individuals at high or low risk of HIV infection. For the first panel, laboratories were free to select type and number of primers; for the second, all were required to use the two primer pairs Pol 3/4 and MMy 9/10' (Nef 1). RESULTS: False-positive and false-negative results were observed in all laboratories (concordance with serology ranged from 40 to 100%). In addition, the number of positive PCR results did not differ significantly between high- and low-risk seronegatives. The use of crude cell lysates in DNA preparation produced the same PCR results as phenol-extracted DNA. Discrepancies between laboratories indicated that factors other than primer pairs contributed strongly to laboratory variability. CONCLUSIONS: Our results emphasize the importance of both positive and negative controls in PCR and demonstrate the value of multicentre PCR quality control.
Authors: B Yen-Lieberman; D Brambilla; B Jackson; J Bremer; R Coombs; M Cronin; S Herman; D Katzenstein; S Leung; H J Lin; P Palumbo; S Rasheed; J Todd; M Vahey; P Reichelderfer Journal: J Clin Microbiol Date: 1996-11 Impact factor: 5.948
Authors: S Hammer; C Crumpacker; R D'Aquila; B Jackson; J Lathey; D Livnat; P Reichelderfer Journal: J Clin Microbiol Date: 1993-10 Impact factor: 5.948
Authors: S Sauvaigo; V Barlet; N Guettari; P Innocenti; F Parmentier; C Bastard; J M Seigneurin; J C Chermann; R Teoule; J Marchand Journal: J Clin Microbiol Date: 1993-05 Impact factor: 5.948
Authors: M G Marin; F Lillo; O E Varnier; S Bresciani; A Molinelli; C Abecasis; P A Bonini; A Albertini Journal: Eur J Clin Microbiol Infect Dis Date: 1995-07 Impact factor: 3.267
Authors: E Papadopulos-Eleopulos; V F Turner; J M Papadimitriou; D Causer; B Hedland-Thomas; B A Page Journal: Genetica Date: 1995 Impact factor: 1.082