Evaluation for the clinical diagnosis of Pythium insidiosum using a single-tube nested PCR.
Pythiosis is a rare infectious disease caused by Pythium insidiosum, which typically occurs in tropical and subtropical regions. The high mortality rate may be in consequence of the lack of diagnosis. The objective of this study was to evaluate reliability of a new single-tube nested PCR for detection of P. insidiosum DNA. A total of 78 clinical isolates of various fungi and bacteria, 106 clinical specimens and 80 simulated positive blood samples were tested. The developed primer pairs CPL6-CPR8 and YTL1-YTR1 are located on 18S subunit of the rRNA gene of P. insidiosum. The specificity, negative and positive predictive values were 100, 100 and 87.5 %, respectively, as compared with direct microscopy and cultivation. The detection limit of the single-tube nested PCR was 21 zoospores corresponding to 2.7 pg of the DNA. The results demonstrate that the new single-tube nested PCR offers a highly sensitive, specific and rapid genetic method for detecting P. insidiosum.
Faculty of Graduated School, Khon Kaen University, Khon Kaen, Thailand.
Yordhathai
Y
Thongsri
Lumyai
L
Wonglakorn
Angkana
A
Chaiprasert
Lucie
L
Svobodova
Petr
P
Hamal
Maitree
M
Pakarasang
Chularut
C
Prariyachatigul
D000293:Adolescent; D000328:Adult; D000368:Aged; D002648:Child; D017931:DNA Primers / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D008828:Microbiological Techniques / Q000379:methods*; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009172:Mycology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D058968:Pythiosis / Q000175:diagnosis* / Q000382:microbiology; D011775:Pythium / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012680:Sensitivity and Specificity; D055815:Young Adult
D017931:DNA Primers; D012337:RNA, Ribosomal, 18S