The rationale and method for constructing internal control DNA used in pertussis polymerase chain reaction
The inclusion of an appropriate internal control DNA in polymerase chain reaction (PCR) is a rapid and simple method for the detection of PCR failure. Two PCR coamplification internal control DNAs (ICD I and ICD II) with the same primer-binding sequences as the target DNA for the detection of Bordet...
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Published in | Diagnostic microbiology and infectious disease Vol. 31; no. 4; pp. 517 - 523 |
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Main Authors | , , , , , |
Format | Journal Article |
Language | English |
Published |
New York, NY
Elsevier Inc
01.08.1998
Elsevier |
Subjects | |
Online Access | Get full text |
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Summary: | The inclusion of an appropriate internal control DNA in polymerase chain reaction (PCR) is a rapid and simple method for the detection of PCR failure. Two PCR coamplification internal control DNAs (ICD I and ICD II) with the same primer-binding sequences as the target DNA for the detection of
Bordetella pertussis and
Bordetella parapertussis were produced using an overlap extension technique and a PCR MIMIC
TM construction kit, respectively. The ICD II was further evaluated in a prospective clinical study in 360 patients with a clinical diagnosis of pertussis. From 360 nasopharyngeal swabs the internal control was positive in 318 (88%) samples, but was negative in 42 (12%). After phenol-chloroform extraction an additional 10 internal controls became positive. For the detection of PCR failure, the use of internal control DNA is highly recommended for PCR-based identification of
B. pertussis and
B. parapertussis organisms from nasopharyngeal swabs and aspirates. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0732-8893 1879-0070 |
DOI: | 10.1016/S0732-8893(98)00043-1 |