The Utility of Multiplex PCR in Simultaneous Detection of HCV, HBV and HIV Infections in Sero-Negative Blood Donors
Mohamed Ahmed Sakr;
Abstract
The risk of transfusion transmitted infections (TTIs) with pathogenic blood-borne viruses such as hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV) has been reduced by improving donor selection and by the development of sensitive serological tests to screen for HBV surface antigen (HBsAg) and antibodies to HCV and HIV. However, a residual risk of viral infection persists related to the pre-seroconversion window period, infection with immunovariant viruses, immunosilent carriage, or occult carriage in the case of HBV infection.
Nucleic acid testing is a molecular technique for screening blood donations to reduce the risk of TTIs in the recipients, thus providing an additional layer of blood safety.
The aim of the current study was to determine the utility of Cobas® TaqScreen Multiplex PCR Test, version 2.0 in simultaneous detection of HCV, HBV and HIV nucleic acids among sero-negative blood donors.
Blood samples were collected from, 5761 blood donors from Blood bank unit of the Central Military Labs for Medical Research. 100 sero-positive blood donor’s samples by using chemiluminescence technique were excluded from the study. The remaining 5661 seronegative samples were divided into 944 pools; each pool was formed of six samples and one pool was formed of 3 samples. Each pool was subjected to the Cobas TaqScreen MPX Test for simultaneous detection of HIV-1 Group M, HIV-1 Group O, HIV-2, HCV-RNA, and HBV-DNA in plasma specimens. Out of 944 pools included in the study, 940 (99.6 %) yielded reliable results and four pools (0.4 %) were repeated due to failure of amplification of internal controls. The percentage of positive pools for HCV-RNA was 0.4% (4/944). Each sample of the four positive pools (24 samples) was tested individually to identify the positive samples. One sample was found to be positive from each of three pools while the fourth positive pool was entirely negative. The percentage of positive samples for HCV was 0.05% (3/5661) and the percentage of false positive results in pools was 0.1%.
The 95% LOD of Cobas® TaqScreen Multiplex PCR for HCV-RNA was estimated by replicate testing using the following dilutions 25 IU/ml, 16 IU/ml and 8 IU/ml, 4 IU/ml and 2 IU/ml of secondary HCV standard and was found to range from 8 to 16 IU/ml.
In conclusion, minipool nucleic acid amplification testing by the Cobas® TaqScreen MPX Test, version 2.0 has helped the blood bank to detect three samples infected with HCV (1: 1887) with a 95% limit of detection between 8 and 16 IU/ml. Failure rate was 0.4% and false positive rate was 0.1%. However
Nucleic acid testing is a molecular technique for screening blood donations to reduce the risk of TTIs in the recipients, thus providing an additional layer of blood safety.
The aim of the current study was to determine the utility of Cobas® TaqScreen Multiplex PCR Test, version 2.0 in simultaneous detection of HCV, HBV and HIV nucleic acids among sero-negative blood donors.
Blood samples were collected from, 5761 blood donors from Blood bank unit of the Central Military Labs for Medical Research. 100 sero-positive blood donor’s samples by using chemiluminescence technique were excluded from the study. The remaining 5661 seronegative samples were divided into 944 pools; each pool was formed of six samples and one pool was formed of 3 samples. Each pool was subjected to the Cobas TaqScreen MPX Test for simultaneous detection of HIV-1 Group M, HIV-1 Group O, HIV-2, HCV-RNA, and HBV-DNA in plasma specimens. Out of 944 pools included in the study, 940 (99.6 %) yielded reliable results and four pools (0.4 %) were repeated due to failure of amplification of internal controls. The percentage of positive pools for HCV-RNA was 0.4% (4/944). Each sample of the four positive pools (24 samples) was tested individually to identify the positive samples. One sample was found to be positive from each of three pools while the fourth positive pool was entirely negative. The percentage of positive samples for HCV was 0.05% (3/5661) and the percentage of false positive results in pools was 0.1%.
The 95% LOD of Cobas® TaqScreen Multiplex PCR for HCV-RNA was estimated by replicate testing using the following dilutions 25 IU/ml, 16 IU/ml and 8 IU/ml, 4 IU/ml and 2 IU/ml of secondary HCV standard and was found to range from 8 to 16 IU/ml.
In conclusion, minipool nucleic acid amplification testing by the Cobas® TaqScreen MPX Test, version 2.0 has helped the blood bank to detect three samples infected with HCV (1: 1887) with a 95% limit of detection between 8 and 16 IU/ml. Failure rate was 0.4% and false positive rate was 0.1%. However
Other data
| Title | The Utility of Multiplex PCR in Simultaneous Detection of HCV, HBV and HIV Infections in Sero-Negative Blood Donors | Other Titles | جدوى استخدام تفاعل البلمرة المتسلسل المتعدد في الكشف المتزامن عن الإصابات بالالتهاب الكبدي الفيروسي سى، بى وفيروس نقص المناعة البشرية في المتبرعين بالدم سلبي المصل | Authors | Mohamed Ahmed Sakr | Issue Date | 2014 |
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