Abstract
Introduction: Giardia lamblia is one of the most prevalent intestinal protozoa in the world, which affect children in both undeveloped and developing countries. This study aimed to determine genotypes of the Giardia lamblia using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP)-PCR techniques. Additionally, the relationship between genotype patterns and their geographical distribution, gender, and age was investigated. Methods: The current study included 926 samples of faeces of children suffering from diarrhoea, who visits the internal clinics at Teaching Hospital, and Child Hospital in AL- Diwaniyah City from November 2012 - Jun 2013. For age groups of 1-12 years exclusively. The samples were examined using a direct mount wet smear, The positive samples were preserved without adding preservatives at a temperature of -20°C until the DNA extraction for G. lamblia genotyping by using PCR and RFLP-PCR technique. Results: Giardia lamblia was detected in 2.15% (20/926) of samples from diarrhea cases in children by amplification of glutamate dehydrogenase gene (gdh) using two specific primers GDHiR and GDHiF. It was revealed that 7 samples belonged to genotype A (35%) and 13 samples belonged to genotype B (65%). All genotype A samples belonged to subgenotype AII (100%), while genotype B samples belonged to subgenotypes BIII (53.61 %) and BIV (47.38 %). Genotype A was detected in children of 1-6 years of age while B genotype was detected in all age groups. Both of the genotypes have been detected in both genders (male and female) and genotype B was found in both urban and rural areas; however, its prevalence was higher in rural areas than in urban areas (100% and 30%, respectively). Conclusion: There are two genotypes of Giardia lamblia, genotype A and genotype B, each of which has secondary genetic patterns which include AII, BIII, and BIV.
Highlights
Giardia lamblia is one of the most prevalent intestinal protozoa in the world, which affect children in both undeveloped and developing countries
Modern molecular techniques such as real-time polymerase chain reaction (PCR) or quantitative PCR are recently used in the diagnosis of certain genes belonging to G. lamblia such as Small subunit ribosomal RNA, these techniques are valuable tools with higher sensitivity compared to conventional PCR in discriminating between strains and genotypes which can be used to understand the molecular epidemiology of G. lamblia [7]
Results of molecular identification of Giardia lamblia using restriction fragment length polymorphism (RFLP)- PCR technique, which is a very sensitive technique for determining genetic variations and distinguishing between subgenotypes, showed the dominance of genotype B with a prevalence of 65% compared with genotype A that reached 35% in Diwaniyah city, this is consistent with a survey [23] conducted in Iran using gdh gene
Summary
Giardia lamblia is one of the most prevalent intestinal protozoa in the world, which affect children in both undeveloped and developing countries. G. duodenalis) is one of the most prevalent intestinal protozoa in the world and one of the most common parasitic causes of Gastroenteritis, which primarily affect children in both undeveloped and developing countries. Boontanom et al revealed that giardiasis infection is prevalent in all age groups, but young children are at the greatest risk for contracting giardiasis especially those attending childcare centers (nurseries) [3]. This parasite is spread throughout the world and it is one of the most common zoonotic agents in humans. Symptoms of G. lamblia infection appear in approximately 200 million people in developing countries located in Asia, Africa, and Latin America [4]
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