title: Assessment of semi-automated nucleic acid testing programme in a Regional Blood Transfusion Centre.
abstract: BACKGROUND
  Detection of human immunodeficiency virus type-1 (HIV-1), hepatitis-C (HCV) and hepatitis-B virus (HBV) in the blood donors is crucial. An efficient form of detection is nucleic acid testing (NAT) in blood screening. We assessed the suitability of commercial NAT testing in a developing country, focusing on the Altona RealStar assay and the method of Sacace Biotechnologies.
  
  
  METHODS
  We have standardised and validated commercially available NAT kits with a semi-automated system for detection of HBV, HCV and HIV-1 in blood donations. The MP-NAT (mini-pool) assay consists of pooling of sample, virus extraction, amplification and detection with commercially available NAT kits. An internal control (IC) is incorporated in the assay to monitor the extraction, target amplification and detection process.
  
  
  RESULTS
  The sensitivity of the Altona RealStar assay at 10-MP for each viral target was evaluated, HBV showed amplification in all diluted positive samples of 100, 50, 25, 10 and 5 IU/ml. HIV and HCV infected samples showed amplification in all diluted positive samples of 500, 100, 50 and 30 IU/ml. For HIV, out of six diluted samples of 30 IU/ml, five were amplified. A total of 14,170 seronegative blood samples were tested by RealStar PCR kit in 10-MP and 6 (0.042%) samples/pools were positive. A total of 65,362 seronegative blood donations were also tested by kits of Sacace Biotechnologies, in 10-MP and 45 (0.075%) pools were positive. The prevalence of combined NAT yield cases among routine donors was 1 in 1559 donations tested for all the 3 viruses.
  
  
  CONCLUSIONS
  The semi-automated combined system for NAT screening assays is robust, sensitive, reproducible, and this gives an additional layer of safety with affordable cost.
authors.affiliation: a Surat Raktadan Kendra & Research Centre , Surat , Gujarat , India.
authors.forename: Kanjaksha
authors.initials: K
authors.lastname: Ghosh
authors.affiliation: a Surat Raktadan Kendra & Research Centre , Surat , Gujarat , India.
authors.forename: Kanchan Kumar
authors.initials: KK
authors.lastname: Mishra
authors.affiliation: a Surat Raktadan Kendra & Research Centre , Surat , Gujarat , India.
authors.forename: Apeksha
authors.initials: A
authors.lastname: Trivedi
authors.affiliation: a Surat Raktadan Kendra & Research Centre , Surat , Gujarat , India.
authors.forename: Sheetal
authors.initials: S
authors.lastname: Sosa
authors.affiliation: a Surat Raktadan Kendra & Research Centre , Surat , Gujarat , India.
authors.forename: Krima
authors.initials: K
authors.lastname: Patel
mesh_terms: D001769:Blood / Q000821:virology*; D001782:Blood Donors; D001803:Blood Transfusion / Q000592:standards*; D015497:HIV-1 / Q000235:genetics*; D006403:Hematologic Tests / Q000295:instrumentation / Q000592:standards*; D006515:Hepatitis B virus / Q000235:genetics*; D006526:Hepatitis C / Q000235:genetics*; D012680:Sensitivity and Specificity
keywords: HBV; HCV; HIV; Nucleic acid testing (NAT); Semi-automated; seronegative