tetano
Editor, Senior Moderator
Clin Chim Acta. 2012 Jun 28. [Epub ahead of print]
A novel high-speed droplet-polymerase chain reaction can detect human influenza virus in less than 30min.
Matsuda K, Yamaguchi A, Taira C, Sueki A, Koeda H, Takagi F, Sugano M, Honda T.
Source
Department of Laboratory Medicine, Shinshu University Hospital, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
Abstract
BACKGROUND:
The polymerase chain reaction (PCR) has been widely used for diagnosis of infection. Rapid detection of influenza virus is useful for therapeutic decisions. We attempted to develop a novel assay by real-time droplet-PCR machine for influenza virus.
METHODS:
RNA extracted from nasal swabs or primary swabs pretreated only were used for PCR analyses. We evaluated reaction time, amplification efficiency, sensitivity, and specificity of the novel droplet-PCR.
RESULTS:
The reaction time of the novel droplet-PCR was 28min, whereas that of PCR using the conventional PCR machine was 80min. The standard curve constructed from the amplification plots by the novel droplet-PCR was: y=-3.6x+42.9; that by PCR using the conventional PCR machine was: y=-3.5x+37.8. The sensitivity and specificity of the novel droplet-PCR were 86.7% and 91.7% for the influenza A and 100.0% and 100.0% for the influenza B, respectively. The novel droplet-PCR provided the specific amplification when using primary swabs without RNA extraction.
CONCLUSIONS:
Our novel droplet-PCR markedly reduced the reaction time while showing same reactivity as that by PCR using the conventional PCR machine. Thus, the novel droplet-PCR assay can be used as a rapid assay for detection of influenza virus.
Copyright ? 2012. Published by Elsevier B.V.
PMID:
22750331
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22750331
A novel high-speed droplet-polymerase chain reaction can detect human influenza virus in less than 30min.
Matsuda K, Yamaguchi A, Taira C, Sueki A, Koeda H, Takagi F, Sugano M, Honda T.
Source
Department of Laboratory Medicine, Shinshu University Hospital, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
Abstract
BACKGROUND:
The polymerase chain reaction (PCR) has been widely used for diagnosis of infection. Rapid detection of influenza virus is useful for therapeutic decisions. We attempted to develop a novel assay by real-time droplet-PCR machine for influenza virus.
METHODS:
RNA extracted from nasal swabs or primary swabs pretreated only were used for PCR analyses. We evaluated reaction time, amplification efficiency, sensitivity, and specificity of the novel droplet-PCR.
RESULTS:
The reaction time of the novel droplet-PCR was 28min, whereas that of PCR using the conventional PCR machine was 80min. The standard curve constructed from the amplification plots by the novel droplet-PCR was: y=-3.6x+42.9; that by PCR using the conventional PCR machine was: y=-3.5x+37.8. The sensitivity and specificity of the novel droplet-PCR were 86.7% and 91.7% for the influenza A and 100.0% and 100.0% for the influenza B, respectively. The novel droplet-PCR provided the specific amplification when using primary swabs without RNA extraction.
CONCLUSIONS:
Our novel droplet-PCR markedly reduced the reaction time while showing same reactivity as that by PCR using the conventional PCR machine. Thus, the novel droplet-PCR assay can be used as a rapid assay for detection of influenza virus.
Copyright ? 2012. Published by Elsevier B.V.
PMID:
22750331
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22750331