The rationale and method for constructing internal control DNA used in pertussis polymerase chain reaction

• Published in: Diagnostic microbiology and infectious disease Vol. 31; no. 4; pp. 517 - 523
• Main Authors: Müller, Frank-Michael C., Schnitzler, Norbert, Cloot, Oliver, Kockelkorn, Peter, Haase, Gerhard, Li, Zhongming
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01.08.1998; Elsevier
• Subjects: Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Bordetella - genetics; Bordetella - isolation & purification; Bordetella pertussis - genetics; Bordetella pertussis - isolation & purification; Child; DNA Primers; Evaluation Studies as Topic; False Negative Reactions; Fundamental and applied biological sciences. Psychology; Humans; Microbiology; Nasopharynx - microbiology; Polymerase Chain Reaction - methods; Prospective Studies; Whooping Cough - diagnosis; Respiratory disease; Bordetella parapertussis; Check; Identification; Bordetella pertussis; Method; Whooping cough; Infection; Polymerase chain reaction; DNA; Bacteriosis; Bacteria; Diagnosis; Detection
• ISSN: 0732-8893; 1879-0070
• DOI: 10.1016/S0732-8893(98)00043-1

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.