Pandemic preparedness in Hawaii: a multicenter verification of real-time RT-PCR for the direct detection of influenza virus types A and B.
OBJECTIVE
We integrated multicenter, real-time (RTi) reverse transcription polymerase chain reaction (RT-PCR) screening into a statewide laboratory algorithm for influenza surveillance and response.


METHODS
Each of three sites developed its own testing strategy and was challenged with one randomized and blinded panel of 50 specimens previously tested for respiratory viruses. Following testing, each participating laboratory reported its results to the Hawaii State Department of Health, State Laboratories Division for evaluation and possible discrepant analysis.


RESULTS
Two of three laboratories reported a 100% sensitivity and specificity, resulting in a 100% positive predictive value and a 100% negative predictive value (NPV) for influenza type A. The third laboratory showed a 71% sensitivity for influenza type A (83% NPV) with 100% specificity. All three laboratories were 100% sensitive and specific for the detection of influenza type B. Discrepant analysis indicated that the lack of sensitivity experienced by the third laboratory may have been due to the analyte-specific reagent probe used by that laboratory. Use of a newer version of the product with a secondary panel of 20 specimens resulted in a sensitivity and specificity of 100%.


CONCLUSIONS
All three laboratories successfully verified their ability to conduct clinical testing for influenza using diverse nucleic acid extraction and RTi RT-PCR platforms. Successful completion of the verification by all collaborating laboratories paved the way for the integration of those facilities into a statewide laboratory algorithm for influenza surveillance and response.
State Laboratories Division, Hawaii State Department of Health, Pearl City, HI 96782, USA. chris.whelen@doh.hawaii.gov
A Christian
AC
Whelen
Matthew J
MJ
Bankowski
Glenn
G
Furuya
Stacey
S
Honda
Robert
R
Ueki
Amelia
A
Chan
Karen
K
Higa
Diane
D
Kumashiro
Nathaniel
N
Moore
Roland
R
Lee
Terrie
T
Koyamatsu
Paul V
PV
Effler
D004279:DNA, Viral / Q000032:analysis; D004196:Disease Outbreaks / Q000517:prevention & control*; D006254:Hawaii / Q000453:epidemiology; D006801:Humans; D009980:Influenza A virus / Q000235:genetics / Q000302:isolation & purification*; D009981:Influenza B virus / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis / Q000453:epidemiology* / Q000517:prevention & control; D007753:Laboratories / Q000592:standards*; D011785:Quality Assurance, Health Care / Q000592:standards*; D011786:Quality Control; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D018571:Sentinel Surveillance
D004279:DNA, Viral