<p>The aim of this study was to investigate the genetic diversity of the SLC22A1 gene and to deduce its possible pharmacogenetic implications within the Cape Coloured population of South&nbsp / Africa / a uniquely admixed population of immigrant Europeans, Asians and the indigenous populations. Recent studies have reported an abundance of polymorphic variants within this solute&nbsp / carrier transporter gene encoding for the organic cation transporter 1, as well as evidence linking these variants to an effect on metformin uptake. This study included establishing baseline&nbsp / frequency distribution of previously reported alleles for 20 SNP variants within the SLC22A1 gene, as well as the development of SNaPshot&reg / and Multiplex AS-PCR genotyping assays, and&nbsp / also exploring the possibility of using High-resolution melt (HRM) analysis as a costeffective alternative for SNP genotyping. Ethics clearance was obtained from the Ethics Committee of the&nbsp / University of the Western Cape. Biological samples in the form of buccal (oral) swabs were collected from 132 unrelated voluntary donors from the Cape Coloured population residing in the&nbsp / Cape Metropolitan area. Two SNaPshot&reg / Multiplex Systems were specifically designed for the study,successfully optimized and used for genotyping. Hundred genetic profiles were then generated for a total of 20 SNP variants on SLC22A1 gene, using this primer extension-based genotyping method that enables multiplexing up 10 SNPs. Population genetics data obtained for&nbsp / the investigated SNPs were analysed using various statistical analysis software. Important population genetic parameters were calculated, and possible pharmacogenetics implications were then discussed. Among others, allelic and genotypic frequencies, as well as linkage disequilibrium were determined and compared with world populations. Minor deviation from Hardy- Weinberg equilibrium was observed in the Cape Coloured population. No significantLinkage Disequilibrium between the investigated SNPs was observed in this population. A Multiplex allele specific &ndash / PCR (MAS-PCR) genotyping&nbsp / system was successfully designed and optimized for the genotyping of 10 SNPs from the SLC22A1. This system, also developed specifically for this study, was made of 2 multiplexes each covering 5 SNPs. It is an inexpensive genotyping assay that allows for efficient discrimination of SNP polymorphisms in one reaction tube with standard PCR conditions. A pilot study was&nbsp / conducted to explore the possibility of using High-resolution melt (HRM) analysis as a cost-effective alternative for SNP genotyping. In addition to genotyping, HRM analysis can be used to scan&nbsp / large numbers of samples for novel genetic variations.&nbsp / </p>

ESBL-producing Enterobacteriaceae are emerging worldwide and they are frequently multi-drug resistant, thus limiting treatment options for infections caused by these pathogens. The overall aim of the thesis was to investigate ESBL-producing Enterobacteriaceae in a Swedish county. First, we developed a molecular method, a multiplex PCR assay for identification of SHV, TEM and CTX-M genes in clinical isolates of Enterobacteriaceae with an ESBL phenotype. From 2002 until the end of 2007 all isolates of ESBL-producing Enterobacteriaceae in Östergötland, Sweden were further investigated. The prevalence of ESBL-producing Enterobacteriaceae was low, &lt;1%, but increasing,while the antibiotic consumption remained unchanged. CTX-M enzymes, particularly CTX-M group 1, dominate in our region as well as in the rest of Europe. Furthermore, we have investigated antimicrobial susceptibility by performing MIC-testing in a large, well-characterized population of CTX-M-producing E. coli. Only three oral antimicrobial agents (fosfomycin, nitrofurantoin and mecillinam) demonstrated susceptibility above 90%. High susceptibility, &gt;90%, was also demonstrated for carbapenems, colistin, tigecycline and amikacin. Sixty-eight per cent of ESBL-producing E. coli was multi-resistant, and the most common multi-resistance pattern was the ESBL phenotype with decreased susceptibility to trimethoprim, trimethoprim-sulfamethoxazole, ciprofloxacin, gentamicin and tobramycin. Isolates belonging to CTX-M group 9 are generally more susceptible to antibiotics than the CTX-M group 1-producing E. coli. Finally, a prospective multicentre case-control study examined the prevalence of ESBL-producing Enterobacteriaceae in faecal samples before and after travel abroad and the risk factors of acquisition. Sixty-eight of 226 travellers (30%) had ESBL-producing Enterobacteriaceae in the faecal flora. The geographical area visited had the highest impact on acquisition, with highest the risk for travellers visiting the Indian subcontinent, followed by Asia and Africa north of the equator. Also, acquisition of ESBL-producing Enterobacteriaceae during travel is associated with abdominal symptoms such as diarrhoea. Age also seemed to affect the risk of acquiring ESBL-producing Enterobacteriaceae, the highest risks were found among travellers ≥ 65 years. This thesis has contributed to increased understanding of the epidemiology of ESBL-producing Enterobacteriaceae and their susceptibility to both beta-lactam and non-beta-lactam agents.

<p>Sexual assault is a significant problem facing the South African society. In this context, efficient but also affordable genotyping systems are needed for positive identification of criminals in incidences of sexual violence. The aim of this study was therefore to develop non-commercial Y-STR genotyping systems suitable for sexual assault cases in South Africa. Y-chromosome STR loci constituting the minimal haplotype are still the most widely used loci in investigating sexual assault cases despite the fact that DYS391 and DYS392 have shown low levels of polymorphism in Xhosa populations in Cape Town. The minimal haplotype was, therefore, further investigated in the Cape Muslim population. The Cape Muslim population generally exhibited high GD values among all the South African populations. These values were higher than 0.5 for most loci, and ranged from 0.447 for DYS391 to 0.957 for DYS385. The highest number of alleles in most loci was also recorded in this population. The overall assessment of the minimal haplotype has shown that this system is still a useful in investigating sexual assault case in many South African subpopulations. Therefore the exercise of internal validation of the minimal haplotype system was successfully carried out in the laboratory. iii The properties of additional novel and widely used STRs were also investigated in this study. Loci were successfully sequenced and allele nomenclature was assigned to them according to the ISFG guidelines.</p>

<p>The aim of this study was to investigate the genetic diversity of the SLC22A1 gene and to deduce its possible pharmacogenetic implications within the Cape Coloured population of South&nbsp / Africa / a uniquely admixed population of immigrant Europeans, Asians and the indigenous populations. Recent studies have reported an abundance of polymorphic variants within this solute&nbsp / carrier transporter gene encoding for the organic cation transporter 1, as well as evidence linking these variants to an effect on metformin uptake. This study included establishing baseline&nbsp / frequency distribution of previously reported alleles for 20 SNP variants within the SLC22A1 gene, as well as the development of SNaPshot&reg / and Multiplex AS-PCR genotyping assays, and&nbsp / also exploring the possibility of using High-resolution melt (HRM) analysis as a costeffective alternative for SNP genotyping. Ethics clearance was obtained from the Ethics Committee of the&nbsp / University of the Western Cape. Biological samples in the form of buccal (oral) swabs were collected from 132 unrelated voluntary donors from the Cape Coloured population residing in the&nbsp / Cape Metropolitan area. Two SNaPshot&reg / Multiplex Systems were specifically designed for the study,successfully optimized and used for genotyping. Hundred genetic profiles were then generated for a total of 20 SNP variants on SLC22A1 gene, using this primer extension-based genotyping method that enables multiplexing up 10 SNPs. Population genetics data obtained for&nbsp / the investigated SNPs were analysed using various statistical analysis software. Important population genetic parameters were calculated, and possible pharmacogenetics implications were then discussed. Among others, allelic and genotypic frequencies, as well as linkage disequilibrium were determined and compared with world populations. Minor deviation from Hardy- Weinberg equilibrium was observed in the Cape Coloured population. No significantLinkage Disequilibrium between the investigated SNPs was observed in this population. A Multiplex allele specific &ndash / PCR (MAS-PCR) genotyping&nbsp / system was successfully designed and optimized for the genotyping of 10 SNPs from the SLC22A1. This system, also developed specifically for this study, was made of 2 multiplexes each covering 5 SNPs. It is an inexpensive genotyping assay that allows for efficient discrimination of SNP polymorphisms in one reaction tube with standard PCR conditions. A pilot study was&nbsp / conducted to explore the possibility of using High-resolution melt (HRM) analysis as a cost-effective alternative for SNP genotyping. In addition to genotyping, HRM analysis can be used to scan&nbsp / large numbers of samples for novel genetic variations.&nbsp / </p>

La presente tesis, titulada “Genes, peoples and languages in Central Africa”, examina los patrones de diversidad genética en poblaciones del oeste de Africa central, más específicamente, poblaciones Bantús y Pigmeas de Gabon y Camerún, dos zonas vitales para la comprensión de la expansión Bantú. Se han analizado más de 800 muestras a nivel del cromosoma Y con el fin de caracterizar genéticamente a estas poblaciones, y establecer la relación genética entre ellas. Los resultados han demostrado que la expansión Bantú homogeneizó el acervo genético de las poblaciones Bantús, eliminando la diversidad pre-Bantú, mientras que diversificó aquel de las poblaciones Pigmeas, introduciendo linajes Bantus. Además, se ha visto que el flujo de linajes paternos parece haber tenido una única dirección: de Bantus a Pigmeos. Estos resultados contrastan con aquellos obtenidos para linajes maternos (DNA mitocondrial) en estas zonas, donde se ha observado un considerable flujo genético de Pigmeos a Bantus, sugiriendo un posible sesgo sexual en la tasa de mestizaje entre poblaciones Bantus y Pigmeas. Un hallazgo interesante es la presencia de un linaje no-africano en estas poblaciones de África subsahariana. / The present thesis titled “ Genes, peoples and languages in Central Africa” examines the genetic diversity patterns in populations from west central Africa, more specifically, in Bantu and Pygmy populations from Gabon and Cameroon, two key areas in the understanding of the Bantu expansion. More than 800 samples have been analysed at the Y chromosome level in order to genetically characterise these populations and establish the genetic relationship between them. The results have shown that the Bantu expansion largely homogenised the gene pool of Bantu populations, erasing the pre-Bantu diversity, while it diversified that of Pygmy groups, introducing Bantu lineages into their gene pool. Furthermore, gene flow of paternal lineages seems to have taken place mainly in one direction; from Bantus to Pygmies. These results contrast with those found in studies of maternal (mtDNA) lineages in these areas, where considerable gene flow from Pygmy to Bantu populations have been observed, suggesting possible sex-biased admixtures rates between Bantu and Pygmy populations. An interesting finding, is the significant presence of a non-African lineage in these sub-Saharan populations.

Introdução: O gênero Acinetobacter sp apresenta considerável heterogeneidade possuindo inúmeras espécies. Atualmente, 23 espécies já foram nomeadas e nove outras espécies já foram descritas. Quatro destas espécies possuem contextos clínicos e epidemiológicos diferentes, no entanto são agrupadas em um complexo denominado Acinetobacter calcoaceticus-Acinetobacter baumannii (ABC) devido à sua similaridade genética e fenotípica. Diversos testes para diferenciação do complexo já foram descritos, porém a maioria não pode ser realizado na rotina laboratorial, pois são caros e laboriosos. Objetivos: Avaliar um método rápido e viável na rotina laboratorial, capaz de diferenciar as espécies do complexo ABC; Determinar a prevalência das diferentes espécies do complexo ABC; Avaliar o perfil de suscetibilidade aos antimicrobianos nas diferentes espécies. Métodos: Foram analisadas 118 amostras de dois hospitais de Porto Alegre-RS através do método Multiplex PCR para o gene gyrB e posteriormente confirmadas pelo padrão-ouro: sequenciamento do 16S-23S ITS. O perfil de suscetibilidade foi realizado através de microdiluição em caldo. Resultados: Das 118 amostras identificadas inicialmente como Acinetobacter sp., a grande maioria dos isolados (106 -89.9%) foram identificados como A. baumannii; mas doze isolados foram identificados como sendo das demais espécies do complexo ABC: 6 (5.1%) A. nosocomialis, 5 (4.2%) A. pittii, e 1 (0.8%) A. genoespécie 10, através da técnica de Multiplex PCR. Todos os resultados foram confirmados por sequenciamento. A. baumannii apresentou um elevado nível (72,6%) de resistência ao imipenem em comparação com as outras espécies seguido da espécie A. nosocomialis que apresentou metade de seus isolados resistentes. Todas as espécies apresentaram baixos índices (inferior a 7,5%) de resistência à Polimixina B e Tigeciclina. Conclusão: O Multiplex PCR para o gene gyrB apresentou resultados fidedignos quando comparados ao padrão-ouro, demonstrando, assim, ser um método confiável para a identificação das espécies do complexo ABC. Outras espécies, além de A. baumannii, ABC podem apresentar percentuais significativos de resistência ao imipenem. / Background: Introduction: The genus Acinetobacter sp presents considerable heterogeneity possessing numerous species. Currently, 23 species have been named and nine other species have been described. Four of these species have different clinical and epidemiological contexts, but are grouped in a complex called Acinetobacter calcoaceticus-Acinetobacter baumannii (ABC) due to their genetic and phenotypic similarity. Several tests for differentiation of the complex have been described, but most can not be performed routinely in the laboratory, they are expensive and laborious. Objectives: To evaluate a fast and feasible in routine laboratory able to differentiate the species of the complex ABC; determine the prevalence of different species of the complex ABC; evaluate the antimicrobial susceptibility profile of the different species. Methods: We analyzed 118 samples from two hospitals in Porto Alegre-RS by the method of Multiplex PCR for gene gyrB and subsequently confirmed by the gold standard: sequencing the 16S-23S ITS. The susceptibility profile was performed by microdilution. Results: Of the 118 samples initially identified as Acinetobacter sp. The great majority of isolates (106 -89.9%) were identified as A. baumannii, but twelve isolates were identified as being from other species of the complex ABC: 6 (5.1%) A. nosocomialis, 5 (4.2%) A. pittii, and 1 (0.8%) A. genospécie 10 by Multiplex PCR technique. All results were confirmed by sequencing. A. baumannii showed a high level (72.6%) of imipenem resistance in comparison with the other species followed by the species A. nosocomialis showed that half of his resistant isolates. All species showed low levels (less than 7.5%) of resistance to Polymyxin B and Tigecycline. Conclusion: Multiplex PCR for gene gyrB results presented totally reliable when compared to the gold standard, demonstrating thus be a safe method for the laboratory. Other species besides A. baumannii, ABC may have significant percentages of resistance to imipenem.

Introdução: O gênero Acinetobacter sp apresenta considerável heterogeneidade possuindo inúmeras espécies. Atualmente, 23 espécies já foram nomeadas e nove outras espécies já foram descritas. Quatro destas espécies possuem contextos clínicos e epidemiológicos diferentes, no entanto são agrupadas em um complexo denominado Acinetobacter calcoaceticus-Acinetobacter baumannii (ABC) devido à sua similaridade genética e fenotípica. Diversos testes para diferenciação do complexo já foram descritos, porém a maioria não pode ser realizado na rotina laboratorial, pois são caros e laboriosos. Objetivos: Avaliar um método rápido e viável na rotina laboratorial, capaz de diferenciar as espécies do complexo ABC; Determinar a prevalência das diferentes espécies do complexo ABC; Avaliar o perfil de suscetibilidade aos antimicrobianos nas diferentes espécies. Métodos: Foram analisadas 118 amostras de dois hospitais de Porto Alegre-RS através do método Multiplex PCR para o gene gyrB e posteriormente confirmadas pelo padrão-ouro: sequenciamento do 16S-23S ITS. O perfil de suscetibilidade foi realizado através de microdiluição em caldo. Resultados: Das 118 amostras identificadas inicialmente como Acinetobacter sp., a grande maioria dos isolados (106 -89.9%) foram identificados como A. baumannii; mas doze isolados foram identificados como sendo das demais espécies do complexo ABC: 6 (5.1%) A. nosocomialis, 5 (4.2%) A. pittii, e 1 (0.8%) A. genoespécie 10, através da técnica de Multiplex PCR. Todos os resultados foram confirmados por sequenciamento. A. baumannii apresentou um elevado nível (72,6%) de resistência ao imipenem em comparação com as outras espécies seguido da espécie A. nosocomialis que apresentou metade de seus isolados resistentes. Todas as espécies apresentaram baixos índices (inferior a 7,5%) de resistência à Polimixina B e Tigeciclina. Conclusão: O Multiplex PCR para o gene gyrB apresentou resultados fidedignos quando comparados ao padrão-ouro, demonstrando, assim, ser um método confiável para a identificação das espécies do complexo ABC. Outras espécies, além de A. baumannii, ABC podem apresentar percentuais significativos de resistência ao imipenem. / Background: Introduction: The genus Acinetobacter sp presents considerable heterogeneity possessing numerous species. Currently, 23 species have been named and nine other species have been described. Four of these species have different clinical and epidemiological contexts, but are grouped in a complex called Acinetobacter calcoaceticus-Acinetobacter baumannii (ABC) due to their genetic and phenotypic similarity. Several tests for differentiation of the complex have been described, but most can not be performed routinely in the laboratory, they are expensive and laborious. Objectives: To evaluate a fast and feasible in routine laboratory able to differentiate the species of the complex ABC; determine the prevalence of different species of the complex ABC; evaluate the antimicrobial susceptibility profile of the different species. Methods: We analyzed 118 samples from two hospitals in Porto Alegre-RS by the method of Multiplex PCR for gene gyrB and subsequently confirmed by the gold standard: sequencing the 16S-23S ITS. The susceptibility profile was performed by microdilution. Results: Of the 118 samples initially identified as Acinetobacter sp. The great majority of isolates (106 -89.9%) were identified as A. baumannii, but twelve isolates were identified as being from other species of the complex ABC: 6 (5.1%) A. nosocomialis, 5 (4.2%) A. pittii, and 1 (0.8%) A. genospécie 10 by Multiplex PCR technique. All results were confirmed by sequencing. A. baumannii showed a high level (72.6%) of imipenem resistance in comparison with the other species followed by the species A. nosocomialis showed that half of his resistant isolates. All species showed low levels (less than 7.5%) of resistance to Polymyxin B and Tigecycline. Conclusion: Multiplex PCR for gene gyrB results presented totally reliable when compared to the gold standard, demonstrating thus be a safe method for the laboratory. Other species besides A. baumannii, ABC may have significant percentages of resistance to imipenem.

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No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Jessica_Silveira_Vitoria.pdf: 6777460 bytes, checksum: a576eea96dc73d5ae068edefa155b085 (MD5) Previous issue date: 2017-07-31 / Sem bolsa / A alimentação escolar deve ofertar alimentos seguros quanto à sua condição higienicossanitária buscando a proteção e promoção da saúde dos escolares. O objetivo deste estudo foi avaliar as condições higienicossanitárias de escolas municipais da cidade de Pelotas - RS por meio da aplicação de uma lista de verificação e de análises microbiológicas do ar ambiental, da superfície de manipulação de alimentos, da água, de preparações alimentícias e das mãos de manipuladores de alimentos. Além disso verificou-se a presença de genes produtores de enterotoxinas estafilocócicas clássicas (A, B, C, D e E) nos isolados de Staphylococcus spp. Foram realizadas visitas em 15 escolas municipais com aplicação da lista de verificação e coleta das amostras para as análises microbiológicas. Realizaram-se contagens de bactérias aeróbias mesófilas e bolores e leveduras nas amostras de ar ambiental, contagens de coliformes termotolerantes e bactérias aeróbias mesófilas em amostras de superfícies de manipulação e contagem de coliformes totais e Escherichia coli em amostras de água. Nas mãos dos manipuladores foram realizadas análises de coliformes termotolerantes e Staphylococcus spp. Nas preparações alimentícias foram realizadas análises de coliformes termotolerantes, Bacillus cereus, Staphylococcus spp. e Salmonella spp. Os isolados de Staphylococcus spp. foram submetidos à técnica de reação em cadeia da polimerase multiplex a fim de verificar a presença dos genes codificadores de enterotoxinas estafilocócicas clássicas. Os resultados obtidos a partir da lista de verificação apontaram alto percentual de não conformidades, sendo que duas das 15 escolas apresentaram "grau de risco sanitário alto" e todas as outras foram classificadas como “situação de risco sanitário regular”. Em relação às análises microbiológicas, a qualidade do ar ambiental foi satisfatória em todas as escolas avaliadas. Entretanto, foram encontradas contagens acima do permitido para bactérias aeróbias mesófilas em superfícies de manipulação de alimentos e coliformes totais e Escherichia coli em amostras de água. As mãos dos manipuladores e as preparações alimentícias apresentaram contagens acima do recomendado para Estafilococos coagulase positiva e coliformes termotolerantes. Em três isolados de Staphylococcus spp. verificou-se a presença de genes codificadores de enterotoxina estafilocócica B. Pode-se concluir que as escolas que fizeram parte da pesquisa não apresentam padrões higienicossanitários adequados, pois foram encontradas inadequações tanto na avaliação pela lista de verificação como nos resultados das análises microbiológicas. / School feeding should provide safe food for their hygienic and sanitary conditions, seeking to protect and promote the health of students. The aim of this study was to evaluate the sanitary conditions of municipal schools in the city of Pelotas - RS, through the application of a checklist and microbiological analyses of air, food handling surface, water, food preparations and hands of food handlers. In addition, the presence of classical EE genes (A, B, C, D and E) in Staphylococcus spp. was verified. Visits were carried out in 15 municipal schools with application of the checklist and sample collection for microbiological analyses. Counts of mesophilic aerobic bacteria and molds and yeasts in the air samples, counts of thermotolerant coliforms and aerobic mesophilic bacteria in samples of manipulation surfaces and counting of coliforms and Escherichia coli in water samples were carried out. In the food handlers’ hands were counts of thermotolerant coliforms and Staphylococcus spp. In the food preparations, counts of thermotolerant coliforms, Bacillus cereus, Staphylococcus spp. and Salmonella spp. were realized. Isolates from Staphylococcus spp. were submitted to the multiplex polymerase chain reaction technique in order to verify the presence of genes coding for classical staphylococcal enterotoxins. The results obtained from the checklist indicated a high percentage of nonconformities, and two of the 15 schools presented a "high sanitary risk situation" and all others were classified as "regular sanitary risk situation". Regarding the microbiological analyses, the air quality was satisfactory in all schools evaluated. However, counts above that allowed for counting of mesophilic aerobic bacteria in food and total coliform surfaces and Escherichia coli in water samples were found. Food handlers’ hands and the food preparations presented counts above the recommended for Staphylococcus coagulase positive and thermotolerant coliforms. In three isolates of Staphylococcus spp. the presence of staphylococcal enterotoxin B encoding genes was verified. It can be concluded that the schools that were part of the research did not present adequate hygienic and sanitary standards, since inadequacies were found both in the evaluation by the checklist and in the results of the microbiological analyses.

Magister Scientiae - MSc / Sexual assault is a significant problem facing the South African society. In this context, efficient but also affordable genotyping systems are needed for positive identification of criminals in incidences of sexual violence. The aim of this study was therefore to develop non-commercial Y-STR genotyping systems suitable for sexual assault cases in South Africa. Y-chromosome STR loci constituting the minimal haplotype are still the most widely used loci in investigating sexual assault cases despite the fact that DYS391 and DYS392 have shown low levels of polymorphism in Xhosa populations in Cape Town. The minimal haplotype was, therefore, further investigated in the Cape Muslim population. The Cape Muslim population generally exhibited high GD values among all the South African populations. These values were higher than 0.5 for most loci, and ranged from 0.447 for DYS391 to 0.957 for DYS385. The highest number of alleles in most loci was also recorded in this population. The overall assessment of the minimal haplotype has shown that this system is still a useful in investigating sexual assault case in many South African subpopulations. Therefore the exercise of internal validation of the minimal haplotype system was successfully carried out in the laboratory. The properties of additional novel and widely used STRs were also investigated in this study. Loci were successfully sequenced and allele nomenclature was assigned to them according to the ISFG guidelines. / South Africa

Magister Scientiae - MSc / The aim of this study was to investigate the genetic diversity of the SLC22A1 gene and to deduce its possible pharmacogenetic implications within the Cape Coloured population of South Africa; a uniquely admixed population of immigrant Europeans, Asians and the indigenous populations. Recent studies have reported an abundance of polymorphic variants within this solute carrier transporter gene encoding for the organic cation transporter 1, as well as evidence linking these variants to an effect on metformin uptake. This study included establishing baseline frequency distribution of previously reported alleles for 20 SNP variants within the SLC22A1 gene, as well as the development of SNaPshot® and Multiplex AS-PCR genotyping assays, and also exploring the possibility of using High-resolution melt (HRM) analysis as a costeffective alternative for SNP genotyping. Ethics clearance was obtained from the Ethics Committee of the University of the Western Cape. Biological samples in the form of buccal (oral) swabs were collected from 132 unrelated voluntary donors from the Cape Coloured population residing in the Cape Metropolitan area. Two SNaPshot® Multiplex Systems were specifically designed for the study,successfully optimized and used for genotyping. Hundred genetic profiles were then generated for a total of 20 SNP variants on SLC22A1 gene, using this primer extension-based genotyping method that enables multiplexing up 10 SNPs. Population genetics data obtained for the investigated SNPs were analysed using various statistical analysis software. Important population genetic parameters were calculated, and possible pharmacogenetics implications were then discussed. Among others, allelic and genotypic frequencies, as well as linkage disequilibrium were determined and compared with world populations. Minor deviation from Hardy- Weinberg equilibrium was observed in the Cape Coloured population. No significantLinkage Disequilibrium between the investigated SNPs was observed in this population. A Multiplex allele specific – PCR (MAS-PCR) genotyping system was successfully designed and optimized for the genotyping of 10 SNPs from the SLC22A1. This system, also developed specifically for this study, was made of 2 multiplexes each covering 5 SNPs. It is an inexpensive genotyping assay that allows for efficient discrimination of SNP polymorphisms in one reaction tube with standard PCR conditions. A pilot study was conducted to explore the possibility of using High-resolution melt (HRM) analysis as a cost-effective alternative for SNP genotyping. In addition to genotyping, HRM analysis can be used to scan large numbers of samples for novel genetic variations. / South Africa