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Development and evaluation of lateral-flow test for rapid detection of Mycoplasma gallisepticum and Mycoplasma synoviae in poultry clinical samples


   Faculty of Health and Life Science

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  Dr Kannan Ganapathy, Dr N Evans  No more applications being accepted  Competition Funded PhD Project (Students Worldwide)

About the Project

Background: Protecting the health of poultry is fundamental in ensuring their welfare, health and production. Often, early diagnosis provides opportunities for early intention to safeguard the health of poultry flocks and avoid production losses. In chicken, Mycoplasma gallisepticum (MG) and Mycoplasmas synoviae (MS) both cause infection and disease affecting the respiratory, reproductive and musculoskeletal systems. Unlike bacteria, mycoplasmas have no cell-wall, is mobile, can survive through harsh environments and resist many antibiotics. Recently, the EU has asserted that the status of MG needs to be declared. The EU has also prohibited the export of eggs/chicks from MG-positive flocks. These vertically transmissible pathogens have been in the poultry industry for many decades and globally cause multibillion-dollar losses. Despite this, there has been no pen-side lateral flow (LF) devices for either detection of antibodies or antigens.

Aims, objectives and approaches: The PhD research focuses on developing and evaluating a lateral flow test for MG and MS antibody/antigen detections in poultry. Single and dual detection LF devices will be developed and validated using laboratory and clinical samples. The objectives to be achieved include (1) synthesis, purification and characterisation of recombinant proteins of MG and MS, (2) production and purification of monoclonal antibodies against mgc2 and vlhA surface proteins of MG and MS respectively, (3) development of procedures for practical running of LF device, including nitrocellulose strip preparation, colloidal-gold conjugation with specific antibodies, and optimisation (4) samples of blood/sera and swabs from a) experimentally infected birds, b) field (farm) infections, (5) standardization of the assay for field use. At each stage, ‘sonicated’ whole antigen proteins will be used for comparison.

The candidate would gain wider experience from working in two different settings, i) UoL and ii) Aviagen UK Limited. At the UoL, training will focus on traditional/molecular bacteriology, biotechnology, LS techniques, designing scientific studies, data analysis and statistics, and scientific writing. The LF test, sensitivity and specificity will be cross-checked with other commonly available laboratory assays (eg. ELISA, culture, PCR, qPCR). Aviagen will provided exposure and training in poultry farms, and at their global laboratory at Edinburgh, Scotland.

The studentship should be commenced before the end of 2022.

HOW TO APPLY

Applications should be made by emailing [Email Address Removed] with:

·        a CV (including contact details of at least two academic (or other relevant) referees);

·         a covering letter – clearly stating your first choice project, and optionally 2nd ranked project, as well as including whatever additional information you feel is pertinent to your application; you may wish to indicate, for example, why you are particularly interested in the selected project(s) and at the selected University;

·        copies of your relevant undergraduate degree transcripts and certificates;

·        a copy of your passport (photo page).

A GUIDE TO THE FORMAT REQUIRED FOR THE APPLICATION DOCUMENTS IS AVAILABLE AT https://www.nld-dtp.org.uk/how-apply. Applications not meeting these criteria may be rejected.

In addition to the above items, please email a completed copy of the Additional Details Form (as a Word document) to [Email Address Removed]. A blank copy of this form can be found at: https://www.nld-dtp.org.uk/how-apply.

Informal enquiries may be made to [Email Address Removed]. The closing date for applications is Friday 8th July 2022 at 12noon (UK time).

Note on English language requirements for international applicants: All students applying to the University of Liverpool must demonstrate that they are competent in the use of the English language and satisfy the University’s requirements. Please see this page for more details on specific requirements: https://www.liverpool.ac.uk/study/international/apply/english-language/.


Funding Notes

CASE studentships are funded by the Biotechnology and Biological Sciences Research Council (BBSRC) for 4 years. Funding will cover tuition fees at the UK rate only, a Research Training and Support Grant (RTSG) and stipend. We aim to support the most outstanding applicants from outside the UK and are able to offer a limited number of bursaries that will enable full studentships to be awarded to international applicants. These full studentships will only be awarded to exceptional quality candidates, due to the competitive nature of this scheme. Note that home (UK) candidates may also apply to this studentship.

References

Influences of swab types and storage temperatures on isolation and molecular detection of Mycoplasma gallisepticum and Mycoplasma synoviae. Avian Pathology, 49, 106-110, 2020.
Co-circulation of genetically diverse population of vaccine related and unrelated respiratory mycoplasmas and viruses in UK poultry flocks with health or production problems. Veterinary Microbiology, 225, 132-138, 2018.

Effects of cyclosporin A on the immune responses and pathogenesis of a virulent strain of Mycoplasma gallisepticum in chickens. Avian Pathology, 32, 495-502, 2003.
Dissecting the molecular diversity and commonality of bovine and human treponemes identifies key survival and adhesion mechanisms. PLoS pathogens. 2021;17(3).
Putative β-Barrel Outer Membrane Proteins of the Bovine Digital Dermatitis-Associated Treponemes: Identification, Functional Characterization, and Immunogenicity Infect Immun. 2020 Apr 20;88(5).
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