Identification of Variable-Number Tandem-Repeat(VNTR)Sequences in Acinetobacter pittii and Development of an Optimized Multiple-Locus VNTR Analysis Typing Scheme

Objective To develop a multiple-locus variable-number tandem-repeat(VNTR)analysis(MLVA)assay for Acinetobacter pittii typing.Methods Polymorphic VNTRs were searched by Tandem Repeats Finder.The distribution and polymorphism of each VNTR locus were analyzed in all the A.pittii genomes deposited in the NCBI genome database by BLAST and were evaluated with a collection of 20 well-characterized clinical A.pittii strains and one reference strain.The MLVA assay was compared with pulsed-field gel electrophoresis(PFGE)for discriminating A.pittii isolates.Results Ten VNTR loci were identified upon bioinformatic screening of A.pittii genomes,but only five of them showed full amplifiability and good polymorphism.Therefore,an MLVA assay composed of five VNTR loci was developed.The typeability,reproducibility,stability,discriminatory power,and epidemiological concordance were excellent.Compared with PFGE,the new optimized MLVA typing scheme provided the same and even greater discrimination.Conclusion Compared with PFGE,MLVA typing is a faster and more standardized alternative for studying the genetic relatedness of A.pittii isolates in disease surveillance and outbreak investigation.