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Simple and specific colorimetric detection of Staphylococcus using its volatile 2-[3-acetoxy-4,4,14-trimethylandrost-8-en-17-yl] propanoic acid in the liquid phase and head space of cultures

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Abstract

Spread of drug-resistant Staphylococcus spp. into communities pose danger demanding effective non-invasive and non-destructive tools for its early detection and surveillance. Characteristic volatile organic compounds (VOCs) produced by bacteria offer new diagnostic targets and novel approaches not exploited so far in infectious disease diagnostics. Our search for such characteristic VOC for Staphylococcus spp. led to the depiction of 2-[3-acetoxy-4,4,14-trimethylandrost-8-en-17-yl] propanoic acid (ATMAP), a moderately volatile compound detected both in the culture and headspace when the organism was grown in tryptone soya broth (TSB) medium. A simple and inexpensive colorimetric method (colour change from yellow to orange) using methyl red as the pH indicator provided an absolutely specific way for identifying Staphylococcus spp., The assay performed in liquid cultures (7-h growth in TSB) as well as in the headspace of plate cultures (grown for 10 h on TSA) was optimised in a 96-well plate and 12-well plate formats, respectively, employing a set of positive and negative strains. Only Staphylococcus spp. showed the distinct colour change from yellow to orange due to the production of the above VOC while in the case of other organisms, the reagent remained yellow. The method validated using known clinical and environmental strains (56 including Staphylococcus, Proteus, Pseudomonas, Klebsiella, Bacillus, Shigella and Escherichia coli) was found to be highly efficient showing 100 % specificity and sensitivity. Such simple methods of bacterial pathogen identification are expected to form the next generation tools for the control of infectious diseases through early detection and surveillance of causative agents.

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Acknowledgments

We are grateful to Mr. Suresh Lingham, M/s Trivitron Pvt Ltd. for clinical samples and Dr. Sridhar, Dept. of Microbiology, Sri Ramachandra University, for providing standard bacterial cultures. Our sincere thanks to P. Dineshkumar and S. Akshay for their contribution to our study. We acknowledge the financial support from Centre with Potential for Excellence in Environmental Science (CPEES) of University Grants Commission. One of the authors, R Aarthi, acknowledges CSIR for providing CSIR-SRF. We also express our deepest gratitude to our family and friends.

Conflict of interest

There is no conflict of interests among the authors for submitting this article. This work was supported by the Centre with Potential for Excellence in Environmental Science (CPEES) of University Grants Commission, India. They have no involvements in the study design; in the collection, analysis and interpretation of data; in the writing of the article and in the decision to submit the article for publication.

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  1. Centre for Biotechnology, Anna University, Sardar Patel road, Guindy, Chennai, 600 025, India

    Raju Saranya, Raju Aarthi & Krishnan Sankaran

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  1. Raju Saranya
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  2. Raju Aarthi
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  3. Krishnan Sankaran
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Correspondence to Krishnan Sankaran.

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Saranya, R., Aarthi, R. & Sankaran, K. Simple and specific colorimetric detection of Staphylococcus using its volatile 2-[3-acetoxy-4,4,14-trimethylandrost-8-en-17-yl] propanoic acid in the liquid phase and head space of cultures. Appl Microbiol Biotechnol 99, 4423–4433 (2015). https://doi.org/10.1007/s00253-015-6573-6

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