Genotyping of <i>Giardia lamblia</i> Human Isolates from Portugal by PCR‐RFLP and Sequencing

Sousa, Maria do Céu; Morais, Joana; Machado, J. E.; Poiares-da-Silva, J.

doi:10.1111/j.1550-7408.2006.00221.x

Cited by 27 publications

(28 citation statements)

References 19 publications

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“…Significant concentrations of the infectious stages of both parasites have been found in water samples collected from rivers in the southern region of Portugal [12,16,17]. Also, studies on human and animal biological reservoirs indicated an important presence of zoonotic species [18][19][20][21][22].…”

Section: Introductionmentioning

confidence: 99%

Presence of Cryptosporidium spp. and Giardia duodenalis in Drinking Water Samples in the North of Portugal

Almeida

Moreira²,

Soares³

et al. 2010

Korean J Parasitol

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Section: Introductionmentioning

confidence: 99%

Presence of Cryptosporidium spp. and Giardia duodenalis in Drinking Water Samples in the North of Portugal

Almeida

Moreira²,

Soares³

et al. 2010

Korean J Parasitol

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“…However, clinical cases among immunocompetent patients are well known [13][14][15][16][17]. Both clinical situations are probably underestimated in the country due to the lack of systematic diagnosis of these parasites.…”

Section: Introductionmentioning

confidence: 99%

Biological and Genetic Characterization of Cryptosporidium spp. and Giardia duodenalis Isolates from Five Hydrographical Basins in Northern Portugal

Almeida¹,

Moreira²,

Soares³

et al. 2010

Korean J Parasitol

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Abstract:To understand the situation of water contamination with Cryptosporidium spp. and Giardia spp. in the northern region of Portugal, we have established a long-term program aimed at pinpointing the sources of surface water and environmental contamination, working with the water-supply industry. Here, we describe the results obtained with raw water samples collected in rivers of the 5 hydrographical basins. A total of 283 samples were analyzed using the Method 1623 EPA, USA. Genetic characterization was performed by PCR and sequencing of genes 18S rRNA of Cryptosporidium spp. and b-giardin of Giardia spp. Infectious stages of the protozoa were detected in 72.8% (206 of 283) of the water samples, with 15.2% (43 of 283) positive for Giardia duodenalis cysts, 9.5% (27 of 283) positive for Cryptosporidium spp. oocysts, and 48.1% (136 of 283) samples positive for both parasites. The most common zoonotic species found were G. duodenalis assemblages A-I, A-II, B, and E genotypes, and Cryptosporidium parvum, Cryptosporidium andersoni, Cryptosporidium hominis, and Cryptosporidium muris. These results suggest that cryptosporidiosis and giardiasis are important public health issues in northern Portugal. To the authors' knowledge, this is the first report evaluating the concentration of environmental stages of Cryptosporidium and Giardia in raw water samples in the northern region of Portugal.

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“…Among these seven assemblages, only A and B have yet been associated with human infections 1,[4][5][6][7][8] , whereas C and D have been identified in dogs, cats, coyotes and wolves, E in cattle, sheep, goats and pigs, and F and G in cats and rats, respectively 1 . Molecular studies have revealed longer genetic distance between these assemblages compared to that used to delineate other protozoal species 9 .…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, it is not known why some patients go asymptomatic while some suffer from clinical symptoms. Host factors as well as differences in the genotypes of Giardia isolates are probably involved in the pathogenesis of the infection 4,10 . There are conflicting reports about the correlation between the genotypes and clinical manifestations of giardiasis; some of them reported an association between mild or intermittent diarrhoea and assemblage A and between severe or persistent diarrhoea and assemblage B 11,12 , while others reported that assemblage B was more prevalent in asymptomatic children [13][14][15] .…”

Section: Introductionmentioning

confidence: 99%

“…Most cases are asymptomatic, but symptomatic infections involving recurrent intestinal symptoms may interrupt the normal growth and development in affected children 2,3 . In addition to its wide spectrum of clinical symptomatology, it also has varied routes of transmission, including anthroponotic, zoonotic, foodborne and waterborne [1][2][3][4] . The conventional diagnostic method for G. lamblia in stool specimens is microscopy, including the examination with wet mount, formalin ethyl acetate concentration and trichrome staining 3 .…”

Section: Introductionmentioning

confidence: 99%

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Bir Türk Hasta Kohortunda Giardia lamblia Genotiplendirilmesi: Semptomlar ile Genotipler Arası İlişkinin Araştırılması

Balcıoğlu¹,

Kurt²,

Sevil³

et al. 2009

Kafkas Univ Vet Fak Derg

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SummaryRecent surveys investigating the molecular biology of Giardia lamblia revealed two distinct assemblages with different clinical outcomes. However, there is not a universal compromise about the clinical effects of each assemblage, warranting further studies. Here, we report the results of the first analyses of the assemblages of G. lamblia in Manisa province located in western Turkey, together with their relationships with the symptoms and DNA sequence analyses of the PCR products. DNA samples were isolated from the stools of 63 patients infected with G. lamblia and 54 DNA samples, amplified successfully with PCR, were digested with the enzyme Xho I for RFLP. Thirty-eight of 54 samples (70.4%) were found to be in Assemblage A, while the remaining 16 samples (29.6%) were found to be in Assemblage B. The number of female patients was found significantly higher in Assemblage B (P=0.18). There was a statistically significant relationship between the occurrence of both abdominal pain and diarrhea and Assemblage B (chi-square, 10.52; P<0.05). No other statistically significant relationship was detected between the assemblages and neither with the symptoms nor with the age groups of the patients. The comparison of the DNA sequences of the PCR products from two assemblage B (one subtype B1 and one B) and one assemblage A samples both with each other and with other DNA sequences in the NCBI website by multialignment analyses, revealed specific regions for assemblages B (B1-B) and A on tpi gene region. Further studies with more patients are required to assess these initial results. Now, our aim is to design a probe for tpi gene region to set up a real-time PCR assay that is easier to conduct and requiring shorter time for the analyses. Keywords: Giardia lamblia, Diagnosis, PCR, Genotyping, SymptomBir Türk Hasta Kohortunda Giardia lamblia Genotiplendirilmesi: Semptomlar ile Genotipler Arası İlişkinin Araştırılması ÖzetGiardia lamblia ile yapılan moleküler düzeydeki çalışmalar iki farklı genotipin farklı klinik belirtilere sahip olduğunu ortaya çıkarmıştır. Bununla birlikte, hangi genotipin ne tür klinik belirtilere neden olduğuna dair henüz kesin bulgular elde edilmiş değildir. Bu araştırmada Türkiye'nin Manisa yöresinden elde edilen G. lamblia izolatları polimeraz zincir reaksiyonu (PZR) ve sonrasında DNA dizi analizi yöntemleriyle araştırılmış ve saptanan genotipler ile hastalardaki klinik semptomlar arasındaki ilişkiler incelenmiştir. Toplam 63 hastadan DNA örnekleri izole edilmiş ve bunların 54'üne Xho I restriksiyon enzimi kullanılarak RFLP analizi uygulanmıştır. Sonuçlar incelendiğinde, 54 örneğin 38'inin (%70.37) A, 16'sının ise (%29.63) B genotipine ait olduğu belirlenmiştir. Kadın hasta sayısının B genotipinde anlamlı düzeyde daha fazla olduğu (p=0.18), ayrıca hastalarda birlikte bulunan karın ağrısı ve ishal yakınması ile G. lamblia B genotipi arasında istatistiksel olarak anlamlı bir ilişki olduğu tespit edilmiştir (ki-kare testi, 10.52; P<0.05). Hastaların yaşı ya da yakınmaları ile G. lamblia genotipleri...

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Genotyping of Giardia lamblia Human Isolates from Portugal by PCR‐RFLP and Sequencing

Cited by 27 publications

References 19 publications

Presence of Cryptosporidium spp. and Giardia duodenalis in Drinking Water Samples in the North of Portugal

Presence of Cryptosporidium spp. and Giardia duodenalis in Drinking Water Samples in the North of Portugal

Biological and Genetic Characterization of Cryptosporidium spp. and Giardia duodenalis Isolates from Five Hydrographical Basins in Northern Portugal

Bir Türk Hasta Kohortunda Giardia lamblia Genotiplendirilmesi: Semptomlar ile Genotipler Arası İlişkinin Araştırılması

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