The First Large‐Scale Nucleic Acid Amplification Testing (NAT) of Donated Blood Using Multiplex Reagent for Simultaneous Detection of HBV, HCV, and HIV‐1 and Significance of NAT for HBV

Ohnuma, Hiroyuki; Tanaka, Takeshi; Yoshikawa, Akira; Murokawa, Hiroyuki; Minegishi, Kiyoshi; Yamanaka, Retsuji; Lizuka, Hisao; Miyamoto, Masaki; Satoh, Shuhei; Nakahira, Seiji; Tomono, Tsugikazu; Murozuka, Takashi; Takeda, Yoshio; Doi, Yasushi; Mine, Hideko; Yokoyama, Shigeki; Hirose, Toru; Nishioka, Kusuya

doi:10.1111/j.1348-0421.2001.tb01300.x

Cited by 37 publications

(27 citation statements)

References 13 publications

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“…Therefore, other factors might involve in the HBsAgϩ/anti-HBsϩ samples. Recently, nucleic acid testing for HBV genome at the blood bank has been suggested as a way to improve the screening assay for HBsAg antigenemia (13). Our results suggest that blood banks should select an HBsAg test kit equipped not only with high sensitivity and specificity but also with the use of configuration by several monoclonal or polyclonal antibodies in order to bind different epitopes within the common "a" determinant.…”

Section: Discussionmentioning

confidence: 62%

Mutation of the “a” Determinant of HBsAg with Discordant HBsAg Diagnostic Kits

Louisirirotchanakul

Kanoksinsombat²,

Theamboonlert³

et al. 2004

Viral Immunology

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Sera from 1003 in- and out-patients were investigated for hepatitis B surface antigen (HBsAg) mutation at Siriraj Hospital, Bangkok, Thailand. Individual samples were screened using two commercial HBsAg kits on automatic machine-based assays set up in parallel. The first kit was a sandwich ELISA kit that used monoclonal capture/monoclonal conjugate and color detection whereas the second was a sandwich MEIA, using monoclonal capture/polyclonal indicator and fluorochrome determination. Six specimens were found discordant by negative EIA and positive MEIA; two specimens of which were detectable of HBV DNA. Three out of four discordant results were confirmed by an anti-HBs neutralization assay. Based on direct sequencing, one HBsAg/anti-HBs sample with multiple mutations in the S gene was found. The mutation included the common glycine to arginine escape mutation at amino acid position 145 of the "a" determinant. The observation should alert the blood bank to the necessity of using screening kits capable of detecting HBV mutant carriers as well as providing verification for the phenomenon of vaccine-escape mutation in Thailand.

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Section: Discussionmentioning

confidence: 62%

Mutation of the “a” Determinant of HBsAg with Discordant HBsAg Diagnostic Kits

Louisirirotchanakul

Kanoksinsombat²,

Theamboonlert³

et al. 2004

Viral Immunology

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“…HBsAg concentration builds up even more slowly. Thus, it is not surprising that cases exist where HBV DNA is already detectable by moderately sensitive DNA amplification techniques, but HBsAg is still undetectable in spite of a normal epitope structure [1,[11][12][13][14]. Surprising is the length of this period in the case reported here.…”

Section: Discussionmentioning

confidence: 75%

Transmission of Hepatitis B Virus 2 Months Prior to Hepatitis Surface Antigen Positivity of Donor Blood

Meisel¹,

Endres²,

Walter³

et al. 2003

Transfus Med Hemother

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An hepatitis surface antigen(HBsAg)-negative donation transmitted hepatitis B virus (HBV) of the very common genotype A/HBsAg subtype adw2 to the recipient of the erythrocyte concentrate. Two months later the donor became borderline HBsAg-positive. The infectious donation was tested retrospectively by real time PCR and contained approximately 2,000 viral genomes/ml, an amount which would have been well detectable by minipool nucleic acid amplification test (NAT) for HBV DNA at the usual detection limit of 1,000 copies/ml. The case shows that the HBsAg-negative, HBV DNA-positive phase may be longer than previously believed and that HBV DNA screening at a sensitivity achievable by minipool NAT testing would have prevented transmission.

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“…According to the evaluations developed in the UK, USA, and Japan, the use of the NAT-methods for testing donor blood has decreased the risk of HBV, HCV, and HIV-1 transmitting by 90%, 95%, and 80%, respectively (3,31,32). Although several commercial systems are available for simultaneous NAT-testing of HIV-1, HBV, and HCV (510), further development of responsive, specific, and reliable multiplex techniques remains an important goal for their wide routine application.…”

Section: Discussionmentioning

confidence: 99%

An Oligonucleotide Microarray for Multiplex Real-Time PCR Identification of HIV-1, HBV, and HCV

et al. 2008

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We describe a novel microarray-based approach for simultaneous identification and quantification of human immunodeficiency virus type 1 (HIV-1) and hepatitis B and C viruses (HBV and HCV) in donor plasma specimens. The method is based on multiplex real-time RT-PCR performed within the microarray hydrogel pads. Double-stranded amplification products are simultaneously detected using nonspecific SYBR Green I dye due to the reaction run in separate pads bearing 5'-immobilized specific primers. Both the sensitivity and specificity of the assay, based on 132 blood specimens analyzed, were 100% (56, 26, and 8 specimens were seropositive to HBV HCV and HIV-1, respectively; 22 were positive to both HIV-1 and HCV and 2 positive to all three viruses; 18 samples were pathogen-negative). The dynamic range of the quantitative analysis covered a six-order interval ranging from 100 to 106 genome equivalents per assay. The 95% detection limits were 14 gEq for HIV-1, 10 gEq (1.7 IU) for HBV, and 15 gEq (7.5 IU) for HCV per assay. The proposed approach is considered to be versatile and could be adapted for simultaneous identification and quantification of numerous genetic targets.

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The First Large‐Scale Nucleic Acid Amplification Testing (NAT) of Donated Blood Using Multiplex Reagent for Simultaneous Detection of HBV, HCV, and HIV‐1 and Significance of NAT for HBV

Cited by 37 publications

References 13 publications

Mutation of the “a” Determinant of HBsAg with Discordant HBsAg Diagnostic Kits

Mutation of the “a” Determinant of HBsAg with Discordant HBsAg Diagnostic Kits

Transmission of Hepatitis B Virus 2 Months Prior to Hepatitis Surface Antigen Positivity of Donor Blood

An Oligonucleotide Microarray for Multiplex Real-Time PCR Identification of HIV-1, HBV, and HCV

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