ANTIMALARIAL SUSCEPTIBILITY AND DRUG RESISTANCE CHARACTERIZATION STUDIES OF PLASMODIUM SPECIES ISOLATES FROM PATIENTS IN SABON-WUSE, NIGER STATE, NIGERIA


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TABLE OF CONTENTS

Title
Abstract
Table of Contents
Abbreviations

CHAPTER ONE
1.0       Introduction
1.1       Background
1.2       Statement of Research Problem
1.3       Justification
1.4       Aim and Objectives
1.4.1 Aim
1.4.2 Specific Objectives
1.4.3 Research Hypothesis
1.5       Limitation of Research

CHAPTER TWO
2.0       Literature Review
2.1       Historical Outline
2.2       The Malaria
2.2.1 Taxonomy
2.2.2 Life Cycle of Human Malaria Parasite
2.3       The Malaria Disease
2.3.1 Epidemiology
2.3.2 Diagnosis of malaria
2.3.3 Economic implication of malaria
2.4       Treatment of Malaria
2.4.1 Classification of antimalarial drugs
2.4.2 Pharmacology of Antimalarial Agents
2.5       Types of Antimalarial Drug Resistance Mechanisms
2.5.1 Mechanism of chloroquine resistance
2.5.2 Mechanism of antifolate resistance
2.5.3 Mechanism of artemisinin compounds resistance
2.6 In vitro tests
2.7       Molecular characterization of antimalarial resistance genes

CHAPTER THREE
3.0       Materials and Methods
3.1       Materials
3.1.1 Equipment
3.1.2 Drugs
3.1.3 Chemical
3.1.4 Others
3.2       Methods
3.2.1 The Study Centre
3.2.2 Retrospective Study
3.2.3 Patient and Study Protocol
3.2.4 Profile of Plasmodium species isolates
3.2.5Performance of the in vitro micro-test
3.2.6 Examination of the Post-culture Blood Slide
3.2.7 Molecular Analysis of Resistant Strains

CHAPTER FOUR
4.0       Results
4.1       Survey of malaria prevalence in Sabon-Wuse
4.1.1 Prevalence of malaria
4.1.2 Drugs prescribed
4.2       Profile of Plasmodium species
4.3 In vitro Assay
4.3.1 Screening of patients
4.3.2 Evaluation of In vitro susceptibility P. falciparum isolates
4.4       Molecular Analysis

CHAPTER FIVE
5.0       Discussion, Summary and Recommendation
5.1       Discussion
5.2       Summary
5.3       Conclusion
5.4       Recommendation
REFERENCES




ABSTRACT


About half of Nigerian population experience at least one episode of malaria per year, resulting in high morbidity and mortality and loss of economic value. Retrospective study of the profile of malarial patients and the antimalarial drugs prescribed at Umaru Musa Yaradua memorial Hospital, Sabon-Wuse, Niger State, Nigeria were undertaken. The Plasmodium species among the malarial patients were also studied. The susceptibility of the predominant species to the prescribed antimalarial drugs was investigated. PCR analysis of the DNA of the resistant Plasmodiuum species isolates against the test antimalarials were studied using Qiagen DNeasy blood and tissue kit method. The retrospective study showed that Artemisinin Combination Therapy (ACT) was the predominantly prescribed drugs while the month of May and October had the highest incidence of malarial infection in the years studied. Plasmodium falciparum (85.7%) was the predominant malaria parasite isolated from malaria patients in Sabon-Wuse, Niger State, Nigeria. Plasmodium malariae accounts for only 14.5% of the malaria parasite observed in the area. Only 5% of the P. falciparum parasites were sensitive in vitro to Chloroquine. PCR analysis of the DNA indicated the presence of Pfcrt-Ra Gene in the study area, which further confirms resistance to Chloroquine. Plasmodium falciparum isolates were observed to be sensitive in vitro to Artesunate-Amodiaquine drug in the study area. The investigation also shows that all the samples analysed had multi antimalarial drug resistant gene Pfcrt/FB (76T). A high percentage (87.5%) of the samples analysed displayed the multidrug resistant gene Pfcrt/FB (76K) at 76 bp. The result of the PCR analysis of the DNA corroborates the observed in vitro susceptibility studies. Thus, there is the need for periodic antimalarial surveillance in order to curb emergence of multi antimalarial drug resistance as observed in Sabon-Wuse.




CHAPTER ONE

1.0            INTRODUCTION

1.1          Background

Malaria is a disease caused by the parasite called Plasmodium species that is transmitted from human-to-human by the female Anopheles mosquito. Malaria is a preventable and treatable disease. If malaria is diagnosed and treated early, the duration of the infection can be considerably shortened, which in turn reduces the risk of complications and death. The word

―malaria‖ comes from 18th century Italian mal meaning "bad" and aria meaning "air". Most likely, the term was first used by Dr. Francisco Torti in Italy, when people thought the disease was caused by foul air in marshy areas (Opeskin, 2009).


Malaria remains a huge burden for human populations living in tropical areas. More than two million malaria cases globally were recorded in 2010, with the heaviest mortality rates in children living in sub-Saharan Africa (WHO, 2011). During the past decade, there was an increase in malaria control interventions and this has resulted in considerable reductions in morbidity and mortality associated with malaria in parts of Africa (O’Meara et al., 2010, Steketee and Campbell, 2010). The WHO recommended malaria control measures include long-lasting insecticidal nets (LLIN), indoor residual spraying programmes (IRS) and access to artemisinin combination therapy (WHO, 2010). Despite these efforts, malaria continues to pose a major public health threat in many African countries (WHO, 2011).


Currently, malaria can be caused by five Plasmodium species which include Plasmodium falciparum, Plasmodium malariae, Plasmodium ovale, Plasmodium vivax and, more recently,

Plasmodium knowlesi (Singh et al., 2004). Plasmodium falciparum is the most prevalent in `Africa and the most pathogenic of these, but in most malaria endemic regions multiple......


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