Prevalence and genotyping of human isolates of Giardia duodenalis from Albania (original) (raw)
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Identification and Assemblage Types of Giardia duodenalis from Patients in Thrace, Turkey
Infectious Diseases and Clinical Microbiology
Objective: Giardiasis is a common disease, and clinical forms can vary based on the assemblage types of the parasite. Detailed information on the subgenotypes may indicate the transmission routes and enlighten the gaps in the epidemiology of the disease. This study aims to reveal the occurrence of giardiasis in Thrace, Turkey, and assemblage types of Giardia duodenalis. Materials and Methods: In total, 573 stool samples taken from the individuals applied to Tekirdağ Central State Hospital in 2009, were examined by wet-mount and zinc sulfate floatation methods. Giardia-positive 26 samples and 64 samples taken from the individuals with gastrointestinal complaints were analyzed by nested PCR-RFLP to differentiate the assemblage types. Sequence analysis was employed for confirmation of assemblage types and subgenotypes. Results: Giardia spp. cysts were detected in 3.66% and 4.54% of the samples with wetmount and zinc sulphate floatation techniques respectively. A total of 27 samples were found positive by nested PCR-RFLP out of 90 samples. Fifteen samples were determined as assemblage A, 2 and 10 samples as B and B/E mix respectively. Sequence analysis showed that the latter assemblage (B/E mix) as A3-B3 mix. Conclusion: Fast identification techniques, namely zinc sulphate flotation can be used for screening stool samples in order to determine Giardia cysts with considerably high sensitivity and specificity. Based on this method, the occurrence rate of giardiasis was found as 4.54% in the studied group. DNA sequencing is necessary to distinguish assemblages and confirm the results of PCR-RFLP.
Report of Giardia assemblages and giardiasis in residents of Guilan province—Iran
Parasitology Research, 2020
Giardia duodenalis is considered a highly diverse organism that infects a variety of mammalian hosts. Giardiasis is a significant public health problem in Iran. The purpose of this study was to investigate the occurrence of Giardia duodenalis (G. lamblia, G. intestinalis) infections in humans residing in the Guilan province of Iran. Stool samples were collected during 12 months from 8356 individuals that had been referred to certain hospitals in the capital city of Rasht in the Guilan province, of which 4126 were males and 4230 were females. The samples were separated into three groups according to patient age: group A 1-9 years old (n = 483); group B 10-19 years old (n = 491); and group C greater than 20 years old (n = 7382). The wet mount technique was performed directly on 8356 fecal samples for microscopy. Samples were examined using a saline and iodine direct smear technique in order to confirm the presence of G. duodenalis. The results indicated that 2.5% (206/8356) of the samples were identified as positive for G. duodenalis. A total of 30% of the infected patients (n = 62) had no symptoms. In symptomatic cases, the most common symptoms (46%, n = 95) were abdominal cramps and bloating. Twenty-four percent of patients (n = 50) had cramps, bloating, nausea, and diarrhea. Sixty positive samples were sent for G. duodenalis genotyping based on the amplification of the gdh gene. Forty-one PCR products were successfully selected and sequenced, where 38 (92.6%) samples were identified as genotype A/subgenotype II and in three samples (7.4%) genotype B/subgenotype IV. Genotype A-II had a dominant prevalence as compared to the genotype B-IV samples that were identified in the study. Based on the samples provided by the regional teaching hospitals and subsequent sample analysis, the authors concluded that assemblage A-II is most likely the most common Giardia subgroup infection in the Guilan region. Assemblages have been reported in both humans and animals; however, further studies need to investigate the role of domestic animals and water reservoirs as potential sources of Giardia infection in the Guilan region.
Genotyping of Giardia intestinalis isolated from people living in Sivas, Turkey
Turkish Journal of Medical Sciences, 2012
The technique of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) genotyping was used to characterise morphologically identical isolates of Giardia intestinalis from human stool samples. Materials and methods: In this study a total of 17 trophozoite samples, obtained either directly from stool samples or after excystation, or by duodenal aspiration, were used. A set of primers was chosen to amplify the different regions of triose phosphate isomerase (tpi) and a segment of the glutamate dehydrogenase (gdh) genes. A single-stranded conformational polymorphism technique was also used in an attempt to discriminate among some subgroups. Results: Only primers of the 683-bp segment of the tpi gene from the trophozoite samples were suitable for obtaining a PCR product. In the total of 17 trophozoite DNAs where the tpi gene segment was amplified, 9 belonged to assemblage A (53%) and 4 to assemblage B (23.5%). It was not possible to identify assemblages for the remaining 4 samples (23.5%). Conclusion: PCR-RFLP tpi gene application was able to discriminate between G. intestinalis assemblage A and B, but not the other subgroups. Since assemblage A is the more prevalent subgroup compared with assemblage B, this subgroup can be said to be responsible for common Giardia infections in Turkey.
Advanced Biomedical Research, 2012
Background: Giardia duodenalis is one of the most prevalent intestinal parasites of human. It also infects a wide range of mammals. Two genotype of G.duodenalis (A and B) were commonly reported among humans with different frequency of distribution in different geographical locations. This work was conducted to discriminate genotypes of Giardia duodenalis human isolates in Isfahan city using PCR-RFLP. This is the first molecular study on human isolates of G.duodenalis in the area. Materials and Methods: Samples were collected from different health centers of Isfahan city during June 2011 and February 2012. From 175 Giardia positive stool samples 67 specimens were selected randomly. Cysts of Giardia positive samples were concentrated by flotation sucrose. Extraction of genomic DNA from trophozoite and cysts was performed using QIAamp Stool Mini kit with a modified protocol. PCR-RFLP method was used to amplify a fragment of 458bp at the glutamate dehydrogenase locus, and restriction enzymes BspLI and RsaI differentiated human genotypes A and B and their subgroups. Results: PCR-RFLP assay of 67 isolates showed 40 (59.7%) isolates as Genotype A group II, 23 (34.32%) samples as Genotype B Group III and two (2.98%) sample as Genotype B group IV. Mixed genotype of (AII and B) was detected only in two isolates (2.98%). Conclusions: PCR-RFLP assay targeting gdh locus is a sensitive tool and discriminates genotypes, sub genotypes and mixed type of G.duodenalis. Results of our study suggest both anthroponotic and zoonotic origins for the infections respectively.
Iranian journal of parasitology, 2013
Giardia duodenalis is one of the most important human enteric parasites throughout the world. Clinical symptoms of this parasite vary from asymptomatic infection to chronic diarrhea. Still it is not clear, whether different types of pathogenesis are due to different strains of organism or to variable host factors. The purpose of this study was to investigate possible correlation of clinical symptoms with assemblages among symptomatic and asymptomatic cases collected from southwest of Iran. Fecal samples were collected from 100 symptomatic and asymptomatic cases, which were positive for G. duodenalis. The samples were subjected to semi-nested PCR and RFLP for gdh gene. Among symptomatic patients, 54% had mixed genotypes AII and BIII, 28% and 18% of samples indicated assemblages BIII and AII, respectively. In contrast, among asymptomatic cases, 64%, 26% and 10%samples had mixed genotypes, BIII and AII assemblages, respectively. Statistical analysis using Chi- Square test showed that t...
Giardia duodenalis is a common gastrointestinal parasite that infects humans and many other mammals. It is most prevalent in many developing and industrialized countries. G. duodenalis is considered to be a complex species. While no morphological distinction among different assemblages exist, it can be genetically differentiated into eight major assemblages: A to H. The aim of this study was to determine the genetic heterogeneity of G. duodenalis in human isolates (a study conducted for the first time in Syria). 40 fecal samples were collected from three different hospitals during the hot summer season of 2014. Extraction of genomic DNA from all Giardia positive samples (based on a microscopic examination) was performed using QIAamp DNA Stool Mini Kit. ˇ-giardin gene was used to differentiate between different Giardia assemblages. The 514 bp fragment was amplified using the Polymerase Chain Reaction method, followed by digestion in HaeIII restriction enzyme. Our result showed that genotype A was more frequent than genotype B, 27/40 (67.5%); 4/40 (10%) respectively. A mixed genotype of A + B was only detected in 9 isolates (22.5%). This is the first molecular study performed on G. duodenalis isolates in Syria in order to discriminate among the different genotypes. Further expanded studies using more genes are needed to detect and identify the Giardia parasite at the level of assemblage and sub-assemblage.
Molecular Characterization of Giardia duodenalis in Children and Adults Sampled in Algeria
Microorganisms
The molecular epidemiology of giardiasis in Africa remains unclear. A study was carried out across four hospitals in Algeria. A total of 119 fecal samples from 55 children, 37 adults, and 27 individuals of undetermined age, all scored positive for intestinal parasites by microscopy, and were screened by real-time PCR for Giardia. Molecular characterization of Giardia was performed by assemblage-specific PCR and PCR targeting the triose phosphate isomerase gene (tpi). Of the 119 samples, 80 (67%) were Giardia-positive by real-time PCR. For 48 moderately-highly real-time PCR-positive samples, tpi genotyping assigned 22 samples to Assemblage A and 26 to Assemblage B. Contrary to Assemblage A, Assemblage B exhibited substantial genetic diversity and allelic heterozygosity. Assemblage-specific PCR proved to be specific for discriminating Assemblage A or B but not as sensitive as tpi genotyping. We confirmed that real-time PCR is more sensitive than microscopy for detecting Giardia in sto...
Iranian Journal of Parasitology
Background: We aimed at genotyping and evaluating the predominance of G. duodenalis assemblages isolated from patients referred to medical laboratories in Khorramabad, Iran from Nov 2015 to Sep 2016. Hence, the development of a costeffective HRM approach to determine genotypes of G. duodenalis based on the triosephosphate isomerase (tpi) gene was examined and the genotyping results with and without diarrhea was compared. Methods: Seventy G. duodenalis positive fecal samples were collected. A microscopic confirmation for the presence of Giardia spp. was performed, cysts of 70 Giardia spp. positive specimens were concentrated using sucrose flotation technique and sucrose solution PCR amplification was performed on 69 of 70 (98.5%) samples, and High Resolution Melting (HRM) analysis was performed using a software. Results: The results showed two distinct genotypes (assemblages A and B) of G. duodenalis but infections with mixture of both assemblages were not detected. The genotypes of ...
Genotyping of Giardia duodenalis in children in upper Egypt using assemblage- specific PCR technique
PLOS ONE, 2020
Giardia duodenalis is a common gastrointestinal protozoan parasite, causing diarrheal illness in humans worldwide. Yet, the distribution of G. duodenalis genotypes among human patients and their clinical relevance remains controversial. This study aimed to detect G. duodenalis in children in Upper Egypt and identify causative genotypes and elucidate a possible correlation between genotype and clinical presentation. One hundred sixty-five children, regardless of symptoms, were tested for giardiasis. Giardia positive stool samples (40/165) were subjected to PCR amplification targeting the tpi gene with positive PCR results in only 35 cases (87.5%). Assemblage-specific amplification of genotypes (A, B, and the zoonotic E strains) revealed predominantly G. duodenalis Assemblage A (45.7%). Assemblage B and mixed A and B infections were detected in 31.4% and 22.8% of children, respectively. Assemblage E was not detected. G. duodenalis assemblage A was dominant in children who complained of diarrhea and abdominal cramps. In contrast, asymptomatic children with positive stool samples display a higher frequency of assemblage B and mixed infections. The study highlights the predominance of Giardia Assemblage A in our study locality. This study is the first for this endemic area to use the copro-PCR technique for diagnosis and genotyping of giardiasis. Study results show the value of simple species-specific primers for genotyping in communities with little access to laboratory resources. Further genetic studies are needed to clarify the association between parasite genetic diversity and patient symptomatology.