Conjugal transfer of co-trimoxazole resistance gene from non-typeable haemophilus influenzae to haemophilus influenzae type B / Siti Yatimah Mohamad

Haemophilus influenzae is one of the pathogens that reside in human nasopharynx. Amongst all the types of H. influenzae, type b (Hib) is the most invasive while non typeable H. influenzae (NTHi) is an opportunist. Co-trimoxazole (SXT) is one of the drugs of choice to treat respiratory infections du...

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Main Author:	Mohamad, Siti Yatimah
Format:	Thesis
Language:	English
Published:	2018
Subjects:	Medical education Medical schools Research Health Communication
Online Access:	https://ir.uitm.edu.my/id/eprint/88631/1/88631.PDF
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id	my-uitm-ir.88631
record_format	uketd_dc
spelling	my-uitm-ir.886312024-01-12T09:32:21Z Conjugal transfer of co-trimoxazole resistance gene from non-typeable haemophilus influenzae to haemophilus influenzae type B / Siti Yatimah Mohamad 2018 Mohamad, Siti Yatimah Medical education. Medical schools. Research Health Communication Haemophilus influenzae is one of the pathogens that reside in human nasopharynx. Amongst all the types of H. influenzae, type b (Hib) is the most invasive while non typeable H. influenzae (NTHi) is an opportunist. Co-trimoxazole (SXT) is one of the drugs of choice to treat respiratory infections due to H. influenzae. However, this antibiotic has lost its potency as there are multiple reports that showed the emergence of SXT-resistant H. influenzae strains worldwide. A previous report in Malaysia showed that many strains of NTHi were resistant to SXT. When both strains ofNTHi and Hib co-exist in the nasopharynx, there is a possibility that SXT-resistant genes from NTHi can be transferred through conjugation. Therefore, this study intents to elucidate the mechanism of transfer of SXT resistance genes from NTHi to Hib when they are in close contact. Three methods of in-vitro mating (filter, liquid and solid) were performed to transfer SXT-resistant genes from NTHi (strain H152) to an SXT-sensitive Hib (strain H582). Attempts were made by varying the donor-to-recipient ratio (1: 1 and 1: 10). However, all three mating methods used failed to produce any transconjugants. In order to understand the reason of the inability of H 152 to transfer the SXT-resistant genes to H582, the DNA of both strains was sent for whole genome re-sequencing. Comparative analysis of amino acid sequences of both strains revealed that H152 contained 11 amino acids mutation in dihydrofolate reductase (DHFR) and insertion of five amino acids in dihydropteroate synthase (DHPS) that contributed to the SXT resistance. In addition, the attachment site attP, the DNA region for site-specific recombination, was present only in H582 but not in H152. Type IV secretory pathway (VirB4) components required for conjugation were only present in H582 and absent in H 152. As a conclusion, NTHi strain H 152 used in this study was not able to transfer the SXT-resistant gene to H582 due to lack of gene components for conjugation. 2018 Thesis https://ir.uitm.edu.my/id/eprint/88631/ https://ir.uitm.edu.my/id/eprint/88631/1/88631.PDF text en public masters Universiti Teknologi MARA (Kampus Sg. Buloh) Faculty of Medicine Mohd. Zain, Zaini
institution	Universiti Teknologi MARA
collection	UiTM Institutional Repository
language	English
advisor	Mohd. Zain, Zaini
topic	Medical education Medical schools Research Health Communication
spellingShingle	Medical education Medical schools Research Health Communication Mohamad, Siti Yatimah Conjugal transfer of co-trimoxazole resistance gene from non-typeable haemophilus influenzae to haemophilus influenzae type B / Siti Yatimah Mohamad
description	Haemophilus influenzae is one of the pathogens that reside in human nasopharynx. Amongst all the types of H. influenzae, type b (Hib) is the most invasive while non typeable H. influenzae (NTHi) is an opportunist. Co-trimoxazole (SXT) is one of the drugs of choice to treat respiratory infections due to H. influenzae. However, this antibiotic has lost its potency as there are multiple reports that showed the emergence of SXT-resistant H. influenzae strains worldwide. A previous report in Malaysia showed that many strains of NTHi were resistant to SXT. When both strains ofNTHi and Hib co-exist in the nasopharynx, there is a possibility that SXT-resistant genes from NTHi can be transferred through conjugation. Therefore, this study intents to elucidate the mechanism of transfer of SXT resistance genes from NTHi to Hib when they are in close contact. Three methods of in-vitro mating (filter, liquid and solid) were performed to transfer SXT-resistant genes from NTHi (strain H152) to an SXT-sensitive Hib (strain H582). Attempts were made by varying the donor-to-recipient ratio (1: 1 and 1: 10). However, all three mating methods used failed to produce any transconjugants. In order to understand the reason of the inability of H 152 to transfer the SXT-resistant genes to H582, the DNA of both strains was sent for whole genome re-sequencing. Comparative analysis of amino acid sequences of both strains revealed that H152 contained 11 amino acids mutation in dihydrofolate reductase (DHFR) and insertion of five amino acids in dihydropteroate synthase (DHPS) that contributed to the SXT resistance. In addition, the attachment site attP, the DNA region for site-specific recombination, was present only in H582 but not in H152. Type IV secretory pathway (VirB4) components required for conjugation were only present in H582 and absent in H 152. As a conclusion, NTHi strain H 152 used in this study was not able to transfer the SXT-resistant gene to H582 due to lack of gene components for conjugation.
format	Thesis
qualification_level	Master's degree
author	Mohamad, Siti Yatimah
author_facet	Mohamad, Siti Yatimah
author_sort	Mohamad, Siti Yatimah
title	Conjugal transfer of co-trimoxazole resistance gene from non-typeable haemophilus influenzae to haemophilus influenzae type B / Siti Yatimah Mohamad
title_short	Conjugal transfer of co-trimoxazole resistance gene from non-typeable haemophilus influenzae to haemophilus influenzae type B / Siti Yatimah Mohamad
title_full	Conjugal transfer of co-trimoxazole resistance gene from non-typeable haemophilus influenzae to haemophilus influenzae type B / Siti Yatimah Mohamad
title_fullStr	Conjugal transfer of co-trimoxazole resistance gene from non-typeable haemophilus influenzae to haemophilus influenzae type B / Siti Yatimah Mohamad
title_full_unstemmed	Conjugal transfer of co-trimoxazole resistance gene from non-typeable haemophilus influenzae to haemophilus influenzae type B / Siti Yatimah Mohamad
title_sort	conjugal transfer of co-trimoxazole resistance gene from non-typeable haemophilus influenzae to haemophilus influenzae type b / siti yatimah mohamad
granting_institution	Universiti Teknologi MARA (Kampus Sg. Buloh)
granting_department	Faculty of Medicine
publishDate	2018
url	https://ir.uitm.edu.my/id/eprint/88631/1/88631.PDF
_version_	1794192135109476352

Conjugal transfer of co-trimoxazole resistance gene from non-typeable haemophilus influenzae to haemophilus influenzae type B / Siti Yatimah Mohamad

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