Direct PCR detection of Burkholderia cepacia complex and identification of its genomovars by using sputum as source of DNA.
We developed a nested PCR assay that detects the recA gene of the Burkholderia cepacia complex in sputum. The product of the first PCR round is also used to identify the genomovar of the pathogen. The protocol achieves high sensitivity and specificity with simple interpretation of genomovar status.
2nd Department of Pediatrics, 2nd Medical School of Charles University, Prague, Czech Republic. pavel.drevinek@Lfmotol.cuni.cz
Pavel
P
Drevínek
Hana
H
Hrbácková
Ondrej
O
Cinek
Jana
J
Bartosová
Otakar
O
Nyc
Alexandr
A
Nemec
Petr
P
Pohunek
D000293:Adolescent; D000328:Adult; D019121:Burkholderia Infections / Q000175:diagnosis / Q000453:epidemiology / Q000382:microbiology; D016956:Burkholderia cepacia / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D002648:Child; D002675:Child, Preschool; D003550:Cystic Fibrosis / Q000453:epidemiology / Q000382:microbiology*; D018153:Czech Republic / Q000453:epidemiology; D004269:DNA, Bacterial / Q000032:analysis*; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D011938:Rec A Recombinases / Q000235:genetics; D012680:Sensitivity and Specificity; D013183:Sputum / Q000737:chemistry*
D004269:DNA, Bacterial; D011938:Rec A Recombinases