tetano
Editor, Senior Moderator
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Jun 10;963C:134-139. doi: 10.1016/j.jchromb.2014.06.004. [Epub ahead of print]
Fast on-site diagnosis of influenza A virus by Palm PCR and portable capillary electrophoresis.
Lim S1, Nan H1, Lee MJ2, Kang SH3.
Author information
Abstract
A method combining Palm polymerase chain reaction (PCR) and portable capillary electrophoresis (CE) was developed for rapid on-site analysis of influenza A (H1N1) virus. The portable CE system was suitable for rapid diagnosis which was able to detect a sample in ∼4min after sample loading, while the 'Palm PCR' system allowed for high-speed nucleic acid amplification in ∼16min. The analysis time from DNA sample to analysis of amplified target DNA molecule was only ∼20min, which was significantly less than slab gel electrophoresis with other commercially available PCR machine. When the 100-bp DNA ladder was separated, the relative standard deviation values (n=5) for the migration times and peak areas of the 100 and 200-bp DNA molecules were 0.26 and 8.9%. The detection limits were 6.3 and 7.2pg/μL, respectively. The combined method was also able to identify two influenza A-associated genes (the HA and NP genes of the novel H1N1 influenza). CE separation was achieved with a sieving matrix of 1% poly(vinylpyrrolidone) (Mr=1,300,000) in 1? TBE buffer (pH 8.45). The combined Palm PCR-portable CE system should provide an improved, fast on-site molecular genetic diagnostic method.
Copyright ? 2014 Elsevier B.V. All rights reserved.
KEYWORDS:
Fast diagnosis; H1N1 influenza; On-site analysis; Palm PCR; Portable CE
PMID:
24956080
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24956080
Fast on-site diagnosis of influenza A virus by Palm PCR and portable capillary electrophoresis.
Lim S1, Nan H1, Lee MJ2, Kang SH3.
Author information
Abstract
A method combining Palm polymerase chain reaction (PCR) and portable capillary electrophoresis (CE) was developed for rapid on-site analysis of influenza A (H1N1) virus. The portable CE system was suitable for rapid diagnosis which was able to detect a sample in ∼4min after sample loading, while the 'Palm PCR' system allowed for high-speed nucleic acid amplification in ∼16min. The analysis time from DNA sample to analysis of amplified target DNA molecule was only ∼20min, which was significantly less than slab gel electrophoresis with other commercially available PCR machine. When the 100-bp DNA ladder was separated, the relative standard deviation values (n=5) for the migration times and peak areas of the 100 and 200-bp DNA molecules were 0.26 and 8.9%. The detection limits were 6.3 and 7.2pg/μL, respectively. The combined method was also able to identify two influenza A-associated genes (the HA and NP genes of the novel H1N1 influenza). CE separation was achieved with a sieving matrix of 1% poly(vinylpyrrolidone) (Mr=1,300,000) in 1? TBE buffer (pH 8.45). The combined Palm PCR-portable CE system should provide an improved, fast on-site molecular genetic diagnostic method.
Copyright ? 2014 Elsevier B.V. All rights reserved.
KEYWORDS:
Fast diagnosis; H1N1 influenza; On-site analysis; Palm PCR; Portable CE
PMID:
24956080
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24956080