Comparison of conventional, nested, and real-time PCR assays for rapid and accurate detection of Vibrio vulnificus.
We conducted a prospective study to target toxR in the blood of patients with skin and soft tissue infections who were admitted to four tertiary hospitals to assess the clinical usefulness of real-time quantitative PCR (Q-PCR) as a diagnostic technique. We performed conventional PCR (C-PCR), nested PCR (N-PCR), and Q-PCR assays and compared the results to those obtained using the "gold standard" of microbiological culture. The lower detection limit for the Q-PCR assay was 5 x 10(0) copies/microl. By use of blood samples of patients with skin and soft tissue infections, the sensitivities of the C-PCR and N-PCR assays against the target toxR gene of V. vulnificus as diagnostic tools were determined to be 45% and 86%, respectively. The C-PCR and N-PCR assays had specificities of 100% and 73%, respectively. When we adopted a crossing-point (cp) cutoff value of <38 cp as a positive result, the Q-PCR assay had 100% sensitivity and specificity. Q-PCR to detect V. vulnificus-specific genes is not only the most sensitive and specific of the techniques but also the most rapid diagnostic method. Therefore, the appropriate application of the Q-PCR assay using blood is useful for the rapid diagnosis and subsequent treatment of V. vulnificus sepsis.
Kim
Hyong Sun
HS
Department of Internal Medicine, Division of Infectious Disease, Chosun University College of Medicine, Gwang-ju, Republic of Korea.
Kim
Dong-Min
DM
Neupane
Ganesh Prasad
GP
Lee
Yu-mi
YM
Yang
Nam-Woong
NW
Jang
Sook Jin
SJ
Jung
Sook-In
SI
Park
Kyung-Hwa
KH
Park
Hae-Ryoung
HR
Lee
Chang Seop
CS
Lee
Sun Hee
SH
D017931:DNA Primers; D015342:DNA Probes; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014735:Vibrio Infections / Q000175:diagnosis* / Q000382:microbiology; D041261:Vibrio vulnificus
D017931:DNA Primers; D015342:DNA Probes