title: Japanese external quality assessment program to standardize HIV-1 drug-resistance testing (JEQS2010 program) using in vitro transcribed RNA as reference material.
abstract: To design appropriate antiretroviral therapy regimens and avoid the emergence of human immunodeficiency virus (HIV)-1 variants with reduced susceptibility to antiretroviral drugs, genotypic drug-resistance testing (HIV genotyping) is strongly recommended. To monitor the quality of HIV genotyping in Japan, we performed an external quality assessment (EQA), named the Japanese external quality assessment program, to standardize HIV genotyping (JEQS). To accurately evaluate the quality of HIV genotyping, we employed as reference material (RM) a well-characterized sample, in vitro transcribed RNA (trRNA) that includes the HIV gag-pol sequence, and created a JEQS2010 panel consisting of three single variant and three mixed trRNA samples. All 11 participating laboratories showed high concordance rates (>96%) for the single variant samples. Eight laboratories also showed good rates of detecting minor variants, but three laboratories failed to detect the variants comprising one-half of the sample. These three laboratories used a common primer that had four internal mismatches to the minor trRNA clone. This program showed the usefulness of trRNA as RM, the high quality of HIV genotyping, and extensive interlaboratory variation in the ability to detect minor variants. These results suggest that improving the quality of HIV genotyping in Japan requires regularly implementing the EQA program and improving the HIV genotyping protocol in each laboratory.
authors.affiliation: 1 Faculty of Health Sciences, Hokkaido University , Sapporo, Japan .
authors.forename: Shigeru
authors.initials: S
authors.lastname: Yoshida
authors.forename: Junko
authors.initials: J
authors.lastname: Hattori
authors.forename: Masakazu
authors.initials: M
authors.lastname: Matsuda
authors.forename: Kiyomi
authors.initials: K
authors.lastname: Okada
authors.forename: Yukumasa
authors.initials: Y
authors.lastname: Kazuyama
authors.forename: Osamu
authors.initials: O
authors.lastname: Hashimoto
authors.forename: Shiro
authors.initials: S
authors.lastname: Ibe
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Fujisawa
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Chiba
authors.forename: Masashi
authors.initials: M
authors.lastname: Tatsumi
authors.forename: Shingo
authors.initials: S
authors.lastname: Kato
authors.forename: Wataru
authors.initials: W
authors.lastname: Sugiura
mesh_terms: D024882:Drug Resistance, Viral; D060005:Genotyping Techniques / Q000592:standards*; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics*; D006801:Humans; D007564:Japan; D059021:Laboratory Proficiency Testing / Q000379:methods / Q000458:organization & administration*; D008826:Microbial Sensitivity Tests / Q000592:standards