Development Of A Rapid Test For Toxocariasis Using Recombinant Toxocara Cati Tes-120 Protein
Toxocariasis is a neglected zoonotic disease with worldwide distribution and has been shown to be especially prevalent among children from socio-economically disadvantaged populations. The causative agents are Toxocara canis and Toxocara cati in which the definitive hosts are dogs and cats, respe...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2020
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| Subjects: | |
| Online Access: | http://eprints.usm.my/55648/1/Siti%20Naqiuyah%20Tan%20Farrizam_Msc%20in%20Molecular%20Medicine_January%202020%20cut.pdf |
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| Summary: | Toxocariasis is a neglected zoonotic disease with worldwide distribution and
has been shown to be especially prevalent among children from socio-economically
disadvantaged populations. The causative agents are Toxocara canis and Toxocara
cati in which the definitive hosts are dogs and cats, respectively. Although it is
acknowledged that human toxocariasis is caused by both T. canis and T. cati, however,
the importance of T. cati as a zoonotic agent of the disease has been under-recognized
since T. cati TES is rarely used for Toxocara serodiagnosis and T. canis TES antigens
are generally thought to be able to detect infections by both species. Thus, it is highly
likely that some cases of T. cati infection may be missed by tests using only T. canis
antigen. Therefore, it is an urgent need for the development of diagnostic tools that use
both T. cati and T. canis recombinant antigens to enable detection of infections caused
by both species. Previously, a T. cati recombinant protein TES-120 was successfully
produced and cloned into a pGEX-4T-1 expression vector which showed good
diagnostic potential in western blot. However, the yield was low and unsuitable for
rapid test development. In this study, the T. cati DNA insert was cloned into a pET32
expression vector and two short peptides tagged variants [five residues each of aspartic
acid (D) and lysine(K)] were designed to increase the chances of obtaining a good
yield of the soluble recombinant protein. All three variants namely TES-120
cati/pET32a, TES-120 5D cati/pET32a, and TES-120 5K cati/pET32a were expressed
and purified. |
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