tetano
Editor, Senior Moderator
J Clin Microbiol. 2011 Feb 2. [Epub ahead of print]
Evaluation of twelve real-time RT-PCR primer-probe sets for the detection of pandemic influenza A/H1N1 2009.
Yang Y, Huang F, Gonzalez R, Wang W, Lu G, Li Y, Vernet G, Jin Q, Wang J.
State Key Laboratory for Molecular Virology and Genetic Engineering, Institute of Pathogen Biology (IPB), Chinese Academy of Medical Sciences & Peking Union Medical College (CAMS&PUMC), Beijing 100730, China; Christophe M?rieux Laboratory, IPB, CAMS-Fondation M?rieux, Beijing 100730, China; Beijing Centre for Disease Control and Prevention, Beijing 100013, China; Fondation M?rieux, 69365 Lyon, France.
Abstract
Real-time reverse transcriptase polymerase chain reaction (rRT-PCR) assays have greatly contributed to the detection, control, and prevention of the pandemic influenza virus A/H1N1 2009. To improve the rRT-PCR assays for detection of pandemic influenza A/H1N1 2009, we evaluated the sensitivity, specificity, and performance of twelve rRT-PCR primer-probe sets [SW (a)-SW (l)] using a panel of virus strains and clinical specimens. These primer-probe sets were derived from published work and designed for detecting the hemagglutinin (HA) or the neuraminidase (NA) gene of the pandemic influenza A/H1N1 2009 virus. A primer-probe set SW (CDC) developed by the Centers for Disease Control and Prevention (US CDC) to target the HA gene of pandemic influenza A/H1N1 2009, was used as a referee method. Our results demonstrated that although all primer-probe sets in this study had as high as 98.4-100% in silico coverage, some of the primer-probe sets had better specificity, sensitivity, and amplification efficiency than others. Two primer-probe sets, SW (h) and SW (l), which target the NA gene of pandemic influenza A/H1N1 2009 virus, were highly sensitive (10(4) copies/reaction), had high detection rates (56/60, P=0.134 and 59/60, P=1.000), and showed ideal specificity compared with SW (CDC). In addition, a cocktail of primer-probe sets targeted to the HA and NA genes displayed higher detection sensitivity than primer-probe sets targeting only HA or NA alone, indicating that for practical applications, a combination of primer-probes targeting HA and NA genes is the best option for the detection of pandemic influenza A/H1N1 2009.
PMID: 21289144 [PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/21289144
Evaluation of twelve real-time RT-PCR primer-probe sets for the detection of pandemic influenza A/H1N1 2009.
Yang Y, Huang F, Gonzalez R, Wang W, Lu G, Li Y, Vernet G, Jin Q, Wang J.
State Key Laboratory for Molecular Virology and Genetic Engineering, Institute of Pathogen Biology (IPB), Chinese Academy of Medical Sciences & Peking Union Medical College (CAMS&PUMC), Beijing 100730, China; Christophe M?rieux Laboratory, IPB, CAMS-Fondation M?rieux, Beijing 100730, China; Beijing Centre for Disease Control and Prevention, Beijing 100013, China; Fondation M?rieux, 69365 Lyon, France.
Abstract
Real-time reverse transcriptase polymerase chain reaction (rRT-PCR) assays have greatly contributed to the detection, control, and prevention of the pandemic influenza virus A/H1N1 2009. To improve the rRT-PCR assays for detection of pandemic influenza A/H1N1 2009, we evaluated the sensitivity, specificity, and performance of twelve rRT-PCR primer-probe sets [SW (a)-SW (l)] using a panel of virus strains and clinical specimens. These primer-probe sets were derived from published work and designed for detecting the hemagglutinin (HA) or the neuraminidase (NA) gene of the pandemic influenza A/H1N1 2009 virus. A primer-probe set SW (CDC) developed by the Centers for Disease Control and Prevention (US CDC) to target the HA gene of pandemic influenza A/H1N1 2009, was used as a referee method. Our results demonstrated that although all primer-probe sets in this study had as high as 98.4-100% in silico coverage, some of the primer-probe sets had better specificity, sensitivity, and amplification efficiency than others. Two primer-probe sets, SW (h) and SW (l), which target the NA gene of pandemic influenza A/H1N1 2009 virus, were highly sensitive (10(4) copies/reaction), had high detection rates (56/60, P=0.134 and 59/60, P=1.000), and showed ideal specificity compared with SW (CDC). In addition, a cocktail of primer-probe sets targeted to the HA and NA genes displayed higher detection sensitivity than primer-probe sets targeting only HA or NA alone, indicating that for practical applications, a combination of primer-probes targeting HA and NA genes is the best option for the detection of pandemic influenza A/H1N1 2009.
PMID: 21289144 [PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/21289144