title: Comparison of conventional, nested, and real-time PCR assays for rapid and accurate detection of Vibrio vulnificus.
abstract: 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.
authors.affiliation: Department of Internal Medicine, Division of Infectious Disease, Chosun University College of Medicine, Gwang-ju, Republic of Korea.
authors.forename: Hyong Sun
authors.initials: HS
authors.lastname: Kim
authors.forename: Dong-Min
authors.initials: DM
authors.lastname: Kim
authors.forename: Ganesh Prasad
authors.initials: GP
authors.lastname: Neupane
authors.forename: Yu-mi
authors.initials: YM
authors.lastname: Lee
authors.forename: Nam-Woong
authors.initials: NW
authors.lastname: Yang
authors.forename: Sook Jin
authors.initials: SJ
authors.lastname: Jang
authors.forename: Sook-In
authors.initials: SI
authors.lastname: Jung
authors.forename: Kyung-Hwa
authors.initials: KH
authors.lastname: Park
authors.forename: Hae-Ryoung
authors.initials: HR
authors.lastname: Park
authors.forename: Chang Seop
authors.initials: CS
authors.lastname: Lee
authors.forename: Sun Hee
authors.initials: SH
authors.lastname: Lee
mesh_terms: 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
chemical_list: D017931:DNA Primers; D015342:DNA Probes