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A Simple Method for Direct Detection and Discrimination of <i>A. baumannii</i>in Tracheal Aspirates without Culture Isolation

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Advances in Infectious Diseases 2020年第2期10卷 148-159页

作者： Cheguri H. Swathi Sudhaharan Sukanya Vemu Lakshmi Kamaraju Saipriya Venkataraman Sritharan Department of Molecular Diagnostics & Biomarkers Global Medical Education & Research Foundation (GMERF) Hyderabad India Nizam’s Institute of Medical Sciences (NIMS) Hyderabad India Kamineni Academy of Medical Sciences and Research Center Hyderabad India Department of Molecular Diagnostics & Biomarkers Gleneagles Global Hospital Hyderabad India

Currently, available phenotyping and commercial methods report A. baumannii only as Acinetobacter calcoaceticus-baumannii complex (ACB) and do not identify individual members of the complex. This is a single blind study aimed to evaluate certain commonly used species-specific genetic markers namely, Intergenic Transcribed Spacer region in 16S rRNA of A. baumannii (ab-its) and gyrB, for identification of ACB members. These molecular targets were first validated on clinical isolates (n = 200) and subsequently on uncultured tracheal aspirates (n = 172). Among the clinical isolates, 183/200 (91.5%) were positive for ab-its. The clinical isolates 17 (17/200) which are failed to amplify in ab-its PCR were subsequently diagnosed by gyrB PCR as A. calcoaceticus (n = 2), A. pitti (n = 6) and A. nosocomialis (n = 9) but not A. baumannii. Among the tracheal aspirates, 62 samples were reported as sterile in Advanced Expert System of VITEK-2, among the remaining 110 samples, 68.1% (75/110) samples contained ab-its target. Twenty-five of the sterile samples (25/62) were found to contain ab-its target sequence. Since, our sample processing method enabled identification of all the species of ACB complex by PCR even in uncultured tracheal aspirates, adaptation of our protocol would enable same day (6 - 8 h) reporting and help the clinician make evidence based therapeutic decision quickly.

关键词： Acinetobacter baumannii ACB Complex ab-its gyrB PCR Tracheal Aspirates

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