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Identification of genus Acinetobacter: Standardization of in-house PCR and its comparison with conventional
Sughosh S Kulkarni1, Radhika Madalgi1, Ganavalli S Ajantha1
1Department of Microbiology, SDM College of Medical Sciences and Hospital, Dharwad, Karnataka, India.
Background:
Acinetobacter is grouped under nonfermenting Gram-negative bacilli. It is increasingly isolated from pathological samples. The ability of this genus to acquire drug resistance and spread in the hospital settings is posing a grave problem in healthcare. Specific treatment protocols are advocated for Acinetobacter infections. Hence, rapid identification and drug susceptibility profiling are critical in the management of these infections.
Aims:
To standardize an in-house polymerase chain reaction (PCR) for identification of genus Acinetobacter and to compare PCR with two protocols for its phenotypic identification.
Methodology:
A total of 96 clinical isolates of Acinetobacter were included in the study. An in-house PCR for genus level identification of Acinetobacter was standardized. All the isolates were phenotypically identified by two protocols. The results of PCR and phenotypic identification protocols were compared.
Results:
The in-house PCR standardized was highly sensitive and specific for the genus Acinetobacter. There was 100% agreement between the phenotypic and molecular identification of the genus. The preliminary identification tests routinely used in clinical laboratories were also in complete agreement with phenotypic and molecular identification.
Conclusion:
The in-house PCR for genus level identification is specific and sensitive. However, it may not be essential for routine identification as the preliminary phenotypic identification tests used in the clinical laboratory reliably identify the genus Acinetobacter.
Insights
A new polymerase chain reaction (PCR) method accurately identifies Acinetobacter, a drug-resistant bacterium. While sensitive and specific, routine phenotypic tests are sufficient for clinical identification.
Area of Science:
- Clinical microbiology
- Molecular diagnostics
- Bacterial identification
Background:
- Acinetobacter, a Gram-negative bacillus, is increasingly isolated from clinical samples.
- Its propensity for drug resistance and hospital-acquired infections poses significant healthcare challenges.
- Rapid identification and susceptibility profiling are crucial for effective treatment of Acinetobacter infections.
Purpose of the Study:
- To standardize an in-house polymerase chain reaction (PCR) assay for genus-level identification of Acinetobacter.
- To compare the diagnostic performance of the developed PCR method against two established phenotypic identification protocols.
Main Methods:
- Standardization of an in-house PCR assay targeting the Acinetobacter genus.
- Phenotypic identification of 96 clinical Acinetobacter isolates using two distinct protocols.
- Comparative analysis of results obtained from PCR and phenotypic methods.
Main Results:
- The in-house PCR assay demonstrated high sensitivity and specificity for Acinetobacter genus identification.
- A perfect 100% concordance was observed between molecular (PCR) and phenotypic identification results.
- Preliminary identification tests commonly employed in clinical laboratories also showed complete agreement with both PCR and phenotypic methods.
Conclusions:
- The developed in-house PCR method is a reliable and accurate tool for genus-level Acinetobacter identification.
- Routine implementation of this PCR may not be necessary, as standard phenotypic methods provide dependable identification.
- Phenotypic identification tests currently used in clinical settings are sufficient for reliable genus-level Acinetobacter detection.
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