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Journal of Clinical Microbiology, September 2008, p. 2992-2998, Vol. 46, No. 9
0095-1137/08/$08.00+0     doi:10.1128/JCM.00027-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Comparison of Conventional, Nested, and Real-Time PCR Assays for Rapid and Accurate Detection of Vibrio vulnificus{triangledown}

Hyong Sun Kim,1 Dong-Min Kim,1* Ganesh Prasad Neupane,1 Yu-mi Lee,1 Nam-Woong Yang,2 Sook Jin Jang,3 Sook-In Jung,4 Kyung-Hwa Park,4 Hae-Ryoung Park,5 Chang Seop Lee,6 and Sun Hee Lee7

Division of Infectious Disease, Department of Internal Medicine,1 Department of Microbiology,2 Department of Laboratory Medicine, Chosun University College of Medicine,3 Gwang-ju, Department of Internal Medicine, Chonnam National University Medical School, Gwang-ju,4 Dental Science Research Institute and BK21 Project for Dental School, Chonnam National University, Gwang-ju,5 Department of Internal Medicine, Chonbuk National University Medical School, Jeonju,6 Department of Internal Medicine, School of Medicine, Pusan National University, Busan Republic of Korea7

Received 7 January 2008/ Returned for modification 25 April 2008/ Accepted 1 July 2008

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 100 copies/µl. 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.

* Corresponding author. Mailing address: Department of Internal Medicine, Chosun University College of Medicine, 588 Seosuk-dong, Dong-gu, Gwang-ju 501-717, Republic of Korea. Phone: 82-62-220-3108. Fax: 82-62-234-9653. E-mail: drongkim{at}chosun.ac.kr

{triangledown} Published ahead of print on 9 July 2008.

Journal of Clinical Microbiology, September 2008, p. 2992-2998, Vol. 46, No. 9
0095-1137/08/$08.00+0     doi:10.1128/JCM.00027-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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