Comparison of one-tube multiplex PCR, automated ribotyping and intergenic spacer (ITS) sequencing for rapid identification of Acinetobacter baumannii

Abstract

Acinetobacter baumannii has emerged as a serious cause of nosocomial infections. Rapid identification of this pathogen is required so that appropriate therapy can be given and outbreaks controlled. This study evaluated a multiplex PCR and an automated ribotyping system for the rapid identification of Acinetobacter baumannii. In total, 22 different reference strains and 138 clinical isolates of Acinetobacter spp., identified by 16S-23S rRNA intergenic spacer (ITS) sequence analysis, were evaluated. All A. baumannii isolates (82 clinical isolates and one reference strain) were identified by the multiplex PCR method (specificity 100%). The sensitivity and specificity of the ribotyping system for identification of A. baumannii were 85.5% (71/83) and 93.5% (72/77), respectively. An additional 100 clinical isolates belonging to the Acinetobacter calcoaceticus-A. baumannii complex were used to compare these two methods for identification of A. baumannii, and this comparison revealed a level of disagreement of 14% (14 isolates). The accuracy of the multiplex PCR was 100%, which was confirmed by sequence analysis of the ITS and recA gene of these isolates. Thus, the multiplex PCR method dramatically increased the efficiency and speed of A. baumannii identification.

Ocular Infection by Multidrug-Resistant Acinetobacter baumannii Causes Eyeball Evisceration-A Case Report

“…After 48 hours, the culture of the vitreous humor revealed the presence of the antibiotic resistant A. baumannii bacteria through automation identification in VITEK 2 compact AST 281, confirmed by molecular biology (Fig. 1) through species-specific primers that amplify a fragment of 208 base pairs of the internal transcribed spacer (ITS) of A. baumannii ribosomal DNA, by the DNA polymerase chain reaction (PCR) (Chen et al, 2007). On day 15(POD), the patient developed lancinating pain, worsening of the ocular condition, loss of light perception, and no response to previous treatments.…”

Genotyping methods for monitoring the epidemic evolution of A. baumannii strains

“…Species from Acinetobacter can be easily differentiated. However, this method is less discriminatory than the PFGE method [19]. Therefore at present, ribotyping has been replaced by the PFGE method.…”

Identification of aac(2')-I Type b aminoglycoside-modifying enzyme genes in resistant Acinetobacter baumannii

“…The results confirmed that all strains were A. baumannii. The primer sequences used for the 16S-23S rDNA A. baumannii-specific PCR amplification were P1: 5'-CATTATCA CGGTAATTAGTG-3' and P2: 5'-AGAGCACTGTGCACTTAAG-3' (Chen et al, 2007). A genetic testing kit was provided by the Wuxi Institute of Cloning Genetic Technologies (Wuxi, China).…”