Detection of influenza virus from throat and pharyngeal swabs with a nested duplex light cycler RT-PCR.
A rapid duplex RT-PCR method was developed using the Roche LightCycler technology for detection of influenza type A and influenza type B viruses. Ninety-seven clinical specimens were analyzed using the Lightcycler method compared with conventional viral culture. Thirty-seven specimens (38.1%) were positive by RT-PCR using matrix protein (MP) primers for influenza A or B virus, compared to thirteen culture positive specimens (13.4%). All culture positive specimens were also positive by RT-PCR. A nested PCR reaction using hemagglutination (HA) gene primers confirmed all of the earlier positive PCR reactions. This LightCycler PCR technique was found to be more sensitive than viral culture for influenza virus detection.
Landstuhl Regional Medical Center, Dept. of Pathology, Microbiology Division, Bldg. 3738, Room 204, Auf dem Kirchberg, Landstuhl 66849, Germany. michael.koenig@lnd.amedd.army.mil
Michael
M
Koenig
Sandra
S
Kosha
Mark
M
Hickman
David
D
Heath
Scott
S
Riddell
Wade
W
Aldous
D015331:Cohort Studies; D005260:Female; D005858:Germany; D006801:Humans; D009980:Influenza A virus / Q000302:isolation & purification; D009981:Influenza B virus / Q000302:isolation & purification; D007251:Influenza, Human / Q000175:diagnosis / Q000821:virology*; D008297:Male; D009975:Orthomyxoviridae / Q000302:isolation & purification*; D010614:Pharynx / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity