Fast on-site diagnosis of influenza A virus by Palm PCR and portable capillary electrophoresis.
J Chromatogr B Analyt Technol Biomed Life Sci
; 963: 134-9, 2014 Jul 15.
Article
in En
| MEDLINE
| ID: mdl-24956080
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 â¼4 min after sample loading, while the 'Palm PCR' system allowed for high-speed nucleic acid amplification in â¼16 min. The analysis time from DNA sample to analysis of amplified target DNA molecule was only â¼20 min, 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 7.2 pg/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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Polymerase Chain Reaction
/
Electrophoresis, Capillary
/
Influenza, Human
/
Influenza A Virus, H1N1 Subtype
Type of study:
Diagnostic_studies
/
Prognostic_studies
Limits:
Humans
Language:
En
Journal:
J Chromatogr B Analyt Technol Biomed Life Sci
Journal subject:
ENGENHARIA BIOMEDICA
Year:
2014
Document type:
Article