Korean J Clin Pathol.  1997 Dec;17(6):1038-1047.

Molecular Typing of Vibrio vulnificus isolates by Random Amplified Polymorphic DNA (RAPD) Analysis

• No authors listed

Abstract

BACKGROUND: RAPD (random amplified polymorphic DNA) analysis uses arbitrary chosen primers in conjunction with the polymerase chain reaction (PCR), providing a means of comparing overall genetic similarity among differ tint strains. This study was undertaken to determine molecular types and genetic similarity among V. vulnificus isolates by RAPD analysis. We compared the results with serotypes of V. vulnificus.
METHODS
A total of 97 V. vulnificus strains including 69 strains from Chonnam University Hospital (CUH, Kwangju, Korea), 13 strains from Wonkwang University Hospital (WUH; Iksan, Korea), 13 strains from Japanese National Institute of Health (JNIH) and two reference strains (ATCC 33815 find ATCC 27562) were analyzed. The identification and serotyping of V. vulnificus were performed by JNIH. Operon (Operon Technologies, USA) 10-mer kits A (1, 2, 4, 5, 7, 9, 12, 17 and 20) were used for RAPD analysis. RAPD results were analyzed by NTSYS-pc (numerical taxonomy system and multivariate analysis system, version 1.51, Applied Biostatistics Inc, USA).
RESULTS
RAPD analysis of 95 V. vulnificus strains revealed four molecular types at distance value 0.5. Type T was most common (60/95) and included 58 strains from CUH. Type I showed further subdivision of 7 subtypes. Type II (23/95) composed of 11 strains from CUH, nine strains from WUH and three from JNIH and two reference strains. Type III strains (6/95) were four WUH strains and two JNIH strains. All strains of type IV (6/95) were from JNIH. The range of genetic similarity values among V. vulnificus isolates was 0.24 to 1.00. The serotypes of 95 strains were 04 (84.2%), 014 (3.2%), 01 (2.1%), 013 (2.1%) and R (2.1%) in order of frequence. The most common 04 serotype strains distributed type I (60 strains), type II (23 strains), type III (6 strains) and IV (6 strains).
CONCLUSIONS
Although the V. vulnificus isolates showed wide range of genetic similarity values, RAPD analysis could separate V. vulnificus into four molecular types, and the isolates from the different hospitals revealed a tendoncy to belong to the different molecular type. There was no specific correlation between molecular types and serotypes of V. vulnificus.

MeSH Terms

Asian Continental Ancestry Group
Biostatistics
Classification
DNA*
Gwangju
Humans
Jeollabuk-do
Jeollanam-do
Molecular Typing*
Multivariate Analysis
Operon
Polymerase Chain Reaction
Serotyping
Vibrio vulnificus*
Vibrio*
DNA