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Application of IgY antibodies against staphylococcal protein A (SpA) of Staphylococcus aureus for detection and prophylactic functions

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Abstract

In the present study, immunoglobulin Y (IgY) antibodies were raised in hens against the surface staphylococcal protein A (SpA) of Staphylococcus aureus. Anti-SpA IgY were tested in vitro for diagnostic applications, bacteriostatic, and biofilm inhibition effects. A specific and sensitive immunocapture PCR (IPCR) was developed to detect S. aureus from food, clinical, and environmental samples. Anti-SpA IgY were used for capturing S. aureus cells from different matrices. Chicken antibodies were chosen over mammalian antibodies based on its inertness to immunoglobulin (Ig)-binding property of SpA protein. No cross-reactivity was encountered with closely related Gram-positive and Gram-negative food pathogens. Inter-assay variation is < 10%. The assay was found suitable for testing on solid and liquid food samples, skin, and nasal swabs. The assay showed limit of detection of ≥ 102 CFU/mL from broth cultures and 102 to 103 CFU/ml from diverse natural samples. This assay overcomes the false positives commonly encountered while using mammalian immunoglobulins (IgG). Anti-SpA IgY antibodies were tested for their bacteriostatic effect on the growth of S. aureus. IgY antibodies at a concentration of 150 μg/ml inhibited the growth of S. aureus completely indicating the potential of IgY antibodies in neutralization of infectious pathogens. Similarly, anti-SpA IgY at MIC50 concentration reduced biofilm formation by ~ 45%. In view of advantages offered by IgY antibodies for specific detection of S. aureus in immunocapture PCR (IPCR) assay and in vitro neutralization potential of S. aureus, we recommend using IgY over conventional IgG of mammals involving S. aureus and its antigens.

Key points

• IPCR with anti-SpA IgY for S. aureus was specific and sensitive for natural samples.

• Anti-SpA IgY at 150 ug/ml displayed growth inhibition of S. aureus strains temporarily.

• Anti-SpA IgY at MIC50 concentrations inhibited the biofilm formation partially.

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Acknowledgments

Authors are thankful to management and administration of VFTSR for providing necessary facilities and infrastructure for carrying out this work. Rohini Krishna Kota thanks Council of Scientific and Industrial Research (CSIR), Govt. of India for awarding Senior Research Fellowship (SRF) [File no.: 09/1253(0002)/2019-EMR-I].

Funding

Prakash Narayana Reddy thanks INSPIRE division of Department of Science and Technology (DST), government of India for granting INSPIRE faculty award and research grant [File No: DST/INSPIRE/04/2017/000565]. Authors also thank Department of Science and Technology for conferring FIST support to Department of Biotechnology of Vignan’s University for 5 years (2015–2019) [File No: SR/FST/LSI-576/2013 (C)].

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Authors and Affiliations

  1. Department of Biotechnology, Vignan’s Foundation for Science, Technology and Research (Deemed to be University), Guntur–Tenali Highway Vadlamudi, Guntur District, Andhra Pradesh, 522 213, India

    Rohini Krishna Kota, Prakash Narayana Reddy & Krupanidhi Sreerama

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  1. Rohini Krishna Kota
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  2. Prakash Narayana Reddy
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  3. Krupanidhi Sreerama
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Contributions

PNR conceived and designed research. RK and PNR conducted experiments. KS monitored the work progress and provided necessary reagents. PNR and RK analyzed data. PNR and KS wrote the manuscript. All authors read and approved the manuscript.

Corresponding author

Correspondence to Prakash Narayana Reddy.

Ethics declarations

Animal experiments were initiated after taking approval from the Institutional Animals Ethical Committee (IAEC) of Vignan’s University. Guidelines issued by Committee for the purpose of Control and Supervision of Experiments on Animals (CPCSEA), Govt. of India, for care and use of animals were followed. All the animal experiments and procedures employed were approved by IAEC.

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The authors declare that they have no conflicts of interest.

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Kota, R.K., Reddy, P.N. & Sreerama, K. Application of IgY antibodies against staphylococcal protein A (SpA) of Staphylococcus aureus for detection and prophylactic functions. Appl Microbiol Biotechnol 104, 9387–9398 (2020). https://doi.org/10.1007/s00253-020-10912-5

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  • DOI: https://doi.org/10.1007/s00253-020-10912-5

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