Hepatitis C virus-polymerase chain reaction minipool testing: 3 years in the largest Swiss blood transfusion service.
ABSTRACT Hepatitis C virus-polymerase chain reaction (HCV-PCR) minipool testing can improve the safety of labile blood products owing to a reduction in the diagnostic preseroconversion window period. In Switzerland, HCV-PCR minipool testing for the release of labile blood components became mandatory in September 1999. In the largest Swiss blood transfusion centre, HCV-PCR minipool testing began in January 1999. This report analyses the performance of the test during a 3-year period: 1 January 1999 to 31 December 2001.
EDTA-blood was collected in either standard tubes or plasma preparation (PPT) tubes from 10 blood transfusion services in Switzerland and then sent to the Blood Transfusion Service SRC Berne. Up to 48 donor samples were pooled overnight using Tecan Genesis RSP 200/8 pipettors. Viral RNA was extracted by using the Qiagen QIAamp 96 viral RNA BioRobot kit on a BioRobot 9604. For PCR amplification and detection of HCV or internal control (IC) sequences, the Roche Cobas Amplicor v2.0 test kit was used. Data management, pool resolution and identification of positive samples were performed using the PMS Software from Tecan.
In the 3-year period from 1 January 1999 to 31 December 2001, 839056 blood donor samples were tested in minipools of up to 48 samples. Thirty-five HCV-PCR-positive donations were identified. Thirty-four samples had antibodies against HCV and were therefore also detected by screening for antibody to HCV (anti-HCV). In October 2001, one seronegative (but PCR-positive) donor was detected.
HCV-PCR minipool testing was successfully introduced in the largest Swiss blood transfusion service. It was shown that the release of HCV-PCR minipool results can be accomplished concurrently with the results of serological analysis. The challenge with a seronegative, but PCR-positive, donor demonstrates that the minipool testing strategy adds additional safety to blood products.
- [Show abstract] [Hide abstract]
ABSTRACT: The prevalence of Mycobacterium avium subsp. paratuberculosis (Map) infection in sheep was estimated by blood PCR. One thousand and five hundred sheep belonging to 150 sheep flocks of Trás-os-Montes e Alto Douro (TMAD, Portugal) were studied. Animals were divided into two groups of five sheep each according their health status: “apparently healthy animals” and “paratuberculosis suspect animals”. Samples were pooled in groups of five. PCR detected Map DNA in 56 (18.7%) pool samples. The overall individual prevalence of Map in this study ranged from 6.4% to 15.4%. Map was found in 20.7% pooled samples from apparently healthy and in 16.7% pooled samples from suspect animals. The aim of the present study was to estimate the prevalence of ovine paratuberculosis using a PCR pool screen approach. The results showed that a simple blood PCR method could be reliably performed on pools of five samples in order to estimate the prevalence of Map infection in the ovine population.Small Ruminant Research 05/2008; 76(3):201-206. · 1.10 Impact Factor
- Japanese Journal of Transfusion and Cell Therapy 01/2007; 53(6):607-612.
- [Show abstract] [Hide abstract]
ABSTRACT: This study was carried out to determine the prevalence of isolated anti-HBc among blood donors in this province and its impact on rejection of collected blood units. Isolated hepatitis B core positivity was found 15% in blood center but in this population we have found no HBV-DNA positivity. We proposed that in order to detect mutant hepatitis B viruses in blood donor population, multi-center studies must be done in this country.Journal of Medical Sciences(Faisalabad) 03/2008;