Human blastocystis subtyping with subtype specific primer developed from unique sequences of SSU rRNA Gene / Nurul 'Izzati Zulkifli

Blastocystis is a single-celled eukaryotic parasite that commonly infects the lower gastrointestinal tract in humans as well as animals. It is genetically diverse whereby 17 subtypes (STs) has been recorded so far. However, ST3 and ST1 are predominant in human stool samples especially in Southeast A...

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Main Author:	Zulkifli, Nurul 'Izzati
Format:	Thesis
Language:	English
Published:	2017
Subjects:	Invertebrates Parasitology Bacteria
Online Access:	https://ir.uitm.edu.my/id/eprint/50362/1/50362.pdf
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id	my-uitm-ir.50362
record_format	uketd_dc
spelling	my-uitm-ir.503622021-09-13T02:25:58Z Human blastocystis subtyping with subtype specific primer developed from unique sequences of SSU rRNA Gene / Nurul 'Izzati Zulkifli 2017 Zulkifli, Nurul 'Izzati Invertebrates Parasitology Bacteria Blastocystis is a single-celled eukaryotic parasite that commonly infects the lower gastrointestinal tract in humans as well as animals. It is genetically diverse whereby 17 subtypes (STs) has been recorded so far. However, ST3 and ST1 are predominant in human stool samples especially in Southeast Asian countries including Malaysia. There is currently a lack of time- and cost-effective detection methods for these STs. Thus, the present study was carried out to develop subtype-specific primers from unique sequences of the SSU rRNA gene of human Blastocystis sp. In this study, 2 sets of primers for both ST1 and ST3 have been developed via PerlPrimer and Oligo Analyser software using sequences acquired from the NCBI database. Next, PCR amplification, followed by gel electrophoresis (1.5% agarose gel, 145 volt, 60 minutes) was performed. The sensitivity of the primers were tested using 10-fold dilutions. The primers were also tested against common enteric bacteria and parasites to ensure its specificity. Each primer pairs successfully amplified the target region of the Blastocystis samples, whereby 138 bp and 233 bp amplicons were acquired for ST1 and ST3, respectively. Furthermore the detection limits of the multiplex PCR were confirmed at 7 ng/μl for the former, and 70 ng/μl for the latter. The primers were also confirmed to be non-specific to the non-Blastocystis samples. Because of the ease of use, specificity and increase in Blastocystis infection in Malaysia, this assay has the potential to greatly improve clinical diagnosis of Blastocystis infection and becoming method of choice. 2017 Thesis https://ir.uitm.edu.my/id/eprint/50362/ https://ir.uitm.edu.my/id/eprint/50362/1/50362.pdf text en public degree Universiti Teknologi MARA ( Kampus Puncak Alam ) Faculty of Health Sciences Tengku Ahmad Basri, Tengku Shahrul Anuar
institution	Universiti Teknologi MARA
collection	UiTM Institutional Repository
language	English
advisor	Tengku Ahmad Basri, Tengku Shahrul Anuar
topic	Invertebrates Parasitology Bacteria
spellingShingle	Invertebrates Parasitology Bacteria Zulkifli, Nurul 'Izzati Human blastocystis subtyping with subtype specific primer developed from unique sequences of SSU rRNA Gene / Nurul 'Izzati Zulkifli
description	Blastocystis is a single-celled eukaryotic parasite that commonly infects the lower gastrointestinal tract in humans as well as animals. It is genetically diverse whereby 17 subtypes (STs) has been recorded so far. However, ST3 and ST1 are predominant in human stool samples especially in Southeast Asian countries including Malaysia. There is currently a lack of time- and cost-effective detection methods for these STs. Thus, the present study was carried out to develop subtype-specific primers from unique sequences of the SSU rRNA gene of human Blastocystis sp. In this study, 2 sets of primers for both ST1 and ST3 have been developed via PerlPrimer and Oligo Analyser software using sequences acquired from the NCBI database. Next, PCR amplification, followed by gel electrophoresis (1.5% agarose gel, 145 volt, 60 minutes) was performed. The sensitivity of the primers were tested using 10-fold dilutions. The primers were also tested against common enteric bacteria and parasites to ensure its specificity. Each primer pairs successfully amplified the target region of the Blastocystis samples, whereby 138 bp and 233 bp amplicons were acquired for ST1 and ST3, respectively. Furthermore the detection limits of the multiplex PCR were confirmed at 7 ng/μl for the former, and 70 ng/μl for the latter. The primers were also confirmed to be non-specific to the non-Blastocystis samples. Because of the ease of use, specificity and increase in Blastocystis infection in Malaysia, this assay has the potential to greatly improve clinical diagnosis of Blastocystis infection and becoming method of choice.
format	Thesis
qualification_level	Bachelor degree
author	Zulkifli, Nurul 'Izzati
author_facet	Zulkifli, Nurul 'Izzati
author_sort	Zulkifli, Nurul 'Izzati
title	Human blastocystis subtyping with subtype specific primer developed from unique sequences of SSU rRNA Gene / Nurul 'Izzati Zulkifli
title_short	Human blastocystis subtyping with subtype specific primer developed from unique sequences of SSU rRNA Gene / Nurul 'Izzati Zulkifli
title_full	Human blastocystis subtyping with subtype specific primer developed from unique sequences of SSU rRNA Gene / Nurul 'Izzati Zulkifli
title_fullStr	Human blastocystis subtyping with subtype specific primer developed from unique sequences of SSU rRNA Gene / Nurul 'Izzati Zulkifli
title_full_unstemmed	Human blastocystis subtyping with subtype specific primer developed from unique sequences of SSU rRNA Gene / Nurul 'Izzati Zulkifli
title_sort	human blastocystis subtyping with subtype specific primer developed from unique sequences of ssu rrna gene / nurul 'izzati zulkifli
granting_institution	Universiti Teknologi MARA ( Kampus Puncak Alam )
granting_department	Faculty of Health Sciences
publishDate	2017
url	https://ir.uitm.edu.my/id/eprint/50362/1/50362.pdf
_version_	1783734830105624576

Human blastocystis subtyping with subtype specific primer developed from unique sequences of SSU rRNA Gene / Nurul 'Izzati Zulkifli

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