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Bacteriology

An Internal Control for Routine Diagnostic PCR: Design, Properties, and Effect on Clinical Performance

Maurice Rosenstraus, Zhuang Wang, Sheng-Yung Chang, David DeBonville, Joanne P. Spadoro
Maurice Rosenstraus
Roche Molecular Systems, Branchburg, New Jersey 08876
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Zhuang Wang
Roche Molecular Systems, Branchburg, New Jersey 08876
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Sheng-Yung Chang
Roche Molecular Systems, Branchburg, New Jersey 08876
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David DeBonville
Roche Molecular Systems, Branchburg, New Jersey 08876
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Joanne P. Spadoro
Roche Molecular Systems, Branchburg, New Jersey 08876
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DOI: 10.1128/JCM.36.1.191-197.1998
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ABSTRACT

We constructed internal controls (ICs) to provide assurance that clinical specimens are successfully amplified and detected. The IC nucleic acids contain primer binding regions identical to those of the target sequence and contain a unique probe binding region that differentiates the IC from amplified target nucleic acid. Because only 20 copies of the IC are introduced into each test sample, a positive IC signal indicates that amplification was sufficient to generate a positive signal from targets present at the limit of test sensitivity. The COBAS AMPLICOR Chlamydia trachomatis, Neisseria gonorrhoeae, Mycobacterium tuberculosis, and human hepatitis C virus tests exhibited inhibition rates ranging from 5 to 9%. Approximately 64% of these inhibitory specimens were not inhibitory when a second aliquot was tested. Because repeatedly inhibitory specimens were not reported as false negative and because additional infected specimens were detected during retesting, test sensitivities were 1 to 6% greater than they would have been if the IC had not been used.

  • Copyright © 1998 American Society for Microbiology
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An Internal Control for Routine Diagnostic PCR: Design, Properties, and Effect on Clinical Performance
Maurice Rosenstraus, Zhuang Wang, Sheng-Yung Chang, David DeBonville, Joanne P. Spadoro
Journal of Clinical Microbiology Jan 1998, 36 (1) 191-197; DOI: 10.1128/JCM.36.1.191-197.1998

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An Internal Control for Routine Diagnostic PCR: Design, Properties, and Effect on Clinical Performance
Maurice Rosenstraus, Zhuang Wang, Sheng-Yung Chang, David DeBonville, Joanne P. Spadoro
Journal of Clinical Microbiology Jan 1998, 36 (1) 191-197; DOI: 10.1128/JCM.36.1.191-197.1998
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KEYWORDS

polymerase chain reaction

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