Quality Assessment of Hepatitis C Virus Nucleic Acid Amplification Methods

Lelie, P. N.; Cuypers, H. T. M.; Drimmelen, A.A.J. van; Quint, Wim

doi:10.1159/000053403

Cited by 15 publications

(26 citation statements)

References 11 publications

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“…The concentration in the VQC HCV RNA genotype 1 standard was determined by calibration against the Eurohep HCV RNA genotype 1 standard in repeated branched DNA assays (Quantiplex 2.0, Chiron, Emeryville, CA). The concentration of 8.76 × 10 6 geq per mL in the VQC standard compared with 3.79 × 10 6 geq per mL in the Eurohep standard was confirmed by limiting dilution data of the VQC proficiency study in 1997 13 …”

Section: Methodsmentioning

confidence: 68%

Sensitivity of HCV RNA and HIV RNA blood screening assays

Lelie¹,

Drimmelen²,

Cuypers³

et al. 2002

Transfusion

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Section: Methodsmentioning

confidence: 68%

Sensitivity of HCV RNA and HIV RNA blood screening assays

Lelie¹,

Drimmelen²,

Cuypers³

et al. 2002

Transfusion

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“…Other authors (3,7,12,15,22) underlined the benefit of the ability to use large sample volumes in the extractor in order to increase the sensitivity of NASBA-and RT-PCR-basedamplification methods for HIV-1 and HCV detection and quantification. Our study confirmed this benefit for the detection of HBV DNA, as the sensitivity of the PCR method was increased at least 10-fold by using 1 ml of a sample in the extraction compared to the use of 100 l. As the extractor is able to process samples up to 2 ml, it should be possible to increase the sensitivity of our method even more.…”

Section: Discussionmentioning

confidence: 99%

“…The NucliSens Extractor (Organon Teknika, Boxtel, The Netherlands) is an automated nucleic acid isolation system based on the guanidinium thiocyanate (GuSCN)-silica extraction method as described by Boom et al (4). Thus far, the system has been validated for the isolation of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) RNAs from human plasma and serum in combination with nucleic acid sequence-based amplification (NASBA)-and reverse transcription (RT)-PCR-based nucleic acid amplification methods (3,6,7,11,12,15,22).…”

mentioning

confidence: 99%

Efficient Extraction of Virus DNA by NucliSens Extractor Allows Sensitive Detection of Hepatitis B Virus by PCR

Gobbers¹,

Oosterlaken²,

Bussel

et al. 2001

J Clin Microbiol

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The NucliSens Extractor is an automated nucleic acid isolation system based on guanidinium thiocyanate (GuSCN)-silica extraction technology. The system has been validated for the isolation of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) RNAs from human samples in combination with nucleic acid sequence-based amplification-and reverse transcription-PCR-based methods. We evaluated the extractor for hepatitis B virus (HBV) DNA extraction from human samples using a noncommercial HBV DNA PCR. Several sample pretreatment procedures in combination with the extractor were compared with the Qiagen extraction method, and the impact of the sample volume used in the extraction on the sensitivity was investigated. Heating of the lysed sample prior to extractor isolation and the use of a large sample volume resulted in highly sensitive detection of HBV DNA. Incubation of a 1-ml sample in GuSCN at 80°C (10 min) and at 37°C (30 min) allowed detection of 4 and 40 HBV genome equivalents/ml, respectively, in standard dilution panels. Sample lysis in GuSCN at room temperature and proteinase K treatment prior to use of the extractor were less efficient procedures. All clinical samples that were PCR positive after Qiagen extraction and/or that were HBsAg positive were also PCR positive after extractor isolation. HBV DNA, HCV RNA, and HIV type 1 RNA were efficiently coextracted from a single sample, allowing reliable detection of viral genomes.

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“…(1) Mycobacterium tuberculosis (22,23) Brought to you by | New York University Bobst Library Technical Services Authenticated Download Date | 6/3/15 5:16 PM (2) Viral diagnosis (hepatitis C virus, human immunodeficiency virus, hepatitis B virus and cytomegalo virus) (24).…”

Section: Quality Control Testing In Amplification Diagnosticsmentioning

confidence: 99%

Standardization and Quality Control of PCR Analyses

Burkardt¹

2000

CCLM

139

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In the very beginning of polymerase chain reaction (PCR) tests entering the field of diagnosis of infectious agents, the introduction of this technology into routine diagnosis was hampered by its frequent tendency to create false-positive results because of contamination. This problem is now widely solved by the introduction of the uracil-N-glycosylase (UNG) anticontamination technology. However, care must still be taken to avoid other sources of producing false positive results. They might additionally derive from human error and/or insufficient PCR amplification and detection protocols. A special case lies in the fact that PCR also amplifies DNA from dead organisms rendering a result diagnostically correct as positive, but clinically as false-positive. In PCR, as in any other diagnostic test, the risk of creating a false-negative result also exists. In such a case, the most probable source besides human error, low target or poor amplification and detection protocols is an inhibition caused by interfering substances in a patient's sample. Strategies to recognize and overcome this issue are discussed in this article. Finally a few results from quality control studies on amplification technologies in the diagnosis of infectious agents are reviewed.

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Quality Assessment of Hepatitis C Virus Nucleic Acid Amplification Methods

Cited by 15 publications

References 11 publications

Sensitivity of HCV RNA and HIV RNA blood screening assays

Sensitivity of HCV RNA and HIV RNA blood screening assays

Efficient Extraction of Virus DNA by NucliSens Extractor Allows Sensitive Detection of Hepatitis B Virus by PCR

Standardization and Quality Control of PCR Analyses

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