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

Yuan HU; Bo Qing LI; Da Zhi JIN; Li Hua HE; Xiao Xia TAO; Jian Zhong ZHANG

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

Author: Yuan HU ¹ ; Bo Qing LI ² ; Da Zhi JIN ³ ; Li Hua HE ¹ ; Xiao Xia TAO ¹ ; Jian Zhong ZHANG ¹
Author Information

1. State Key Laboratory of Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
2. Binzhou Medical College, Yantai 256603, Shandong, China.
3. Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310009, Zhejiang, China.
Publication Type:Journal Article
Keywords: A. pittii; MLVA typing; VNTR
MeSH: Acinetobacter; classification; genetics; DNA Fingerprinting; methods; Electrophoresis, Gel, Pulsed-Field; Minisatellite Repeats; Polymerase Chain Reaction
From: Biomedical and Environmental Sciences 2015;28(12):855-863
CountryChina
Language:English
Abstract:
OBJECTIVETo develop a multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) assay for Acinetobacter pittii typing.
METHODSPolymorphic 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.
RESULTSTen 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.
CONCLUSIONCompared 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.