Development of HRM real-time PCR for assemblage characterization of Giardia lamblia

Bahramdoost, Zahra; Mirjalali, Hamed; Yavari, Parvin; Haghighi, Ali

doi:10.1016/j.actatropica.2021.106109

Cited by 10 publications

(5 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Because dogs and cats excrete genotypes common to humans, there is a risk of zoonotic transmission, particularly in asymptomatic infections, when carrier animals may excrete cysts into the external environment for a considerable time [ 13 , 27 , 28 , 38 – 40 ]. Therefore, routine testing for G. duodenalis infection in different habitats is of great importance [ 30 ].…”

Section: Discussionmentioning

confidence: 99%

Prevalence of Giardia duodenalis in dogs and cats: Age-related predisposition, symptomatic, and asymptomatic cyst shedding

Kurnosova,

Panova,

Arisov

2024

Vet World

View full text Add to dashboard Cite

Background and Aim: Giardiasis is a protozoal disease that is globally prevalent in dogs and cats. The clinical manifestations vary, but asymptomatic cases have also been reported. This study aimed to estimate the prevalence of Giardia duodenalis in domestic dogs and cats, characterize the age susceptibility to this disease, and determine the frequency of cases in which cysts are combined with stool changes. Materials and Methods: After centrifugation in a zinc sulfate solution (specific gravity = 1.32 g/cm3), feces of dogs (2761) and cats (1579) were examined microscopically. The age of the animals and the presence of coprological stool characteristics such as fecal odor, consistency, presence of mucus, and other pathological impurities were taken into account. Results: G. duodenalis infection rates were 18.2% (215/1182) in dogs aged 1–12 months and 3.8% (60/1579) in dogs older than 12 months. The infection rate was 7.8% (48/615) in cats aged 1–12 months and 3.35% (33/994) in cats aged >12 months. The most frequently observed coprological abnormalities in cyst-positive animals were soft and/or mushy stool and pungent odor. In dogs, the frequency of these symptoms was 24.4% (67/275), 27.6% (76/275), and 36.4% (100/275) for soft stools, mushy stools, and pungent fecal odor, and 37.8% (31/82), 25.6% (21/82), and 19.5% (16/82), respectively. No stool changes were found when G. duodenalis cysts were detected in dogs in 24.7% (68/275) of cases and in cats in 24.4% (20/82). Conclusion: G. duodenalis cysts are reported more frequently in domestic cats and dogs under 12 months of age than in dogs under 12 months of age. The presence of Giardia cysts is usually accompanied by a strong odor of feces and changes in their consistency. It can be concluded that it is necessary to conduct periodic surveillance for apparently healthy dogs and cats to rule out G. duodenalis infection. Keywords: age of animals, cat feces, consistency of dog feces, coprological parameters, cysts, feces, Giardia duodenalis.

show abstract

Section: Discussionmentioning

confidence: 99%

Prevalence of Giardia duodenalis in dogs and cats: Age-related predisposition, symptomatic, and asymptomatic cyst shedding

Kurnosova,

Panova,

Arisov

2024

Vet World

View full text Add to dashboard Cite

show abstract

“…Assemblage A is the predominant genetic lineage of G . duodenalis in humans in most of molecular studies in Iran [ 28 , 34 , 35 ]. In a study conducted by Sarkari et al [ 36 ], assemblage A was identified among 74.4% of Giardia -positive samples based on the amplification followed by restriction fragment length polymorphism (RFLP) of partial fragment of gdh gene.…”

Section: Discussionmentioning

confidence: 99%

“…Mahmoudi et al [28] reported assemblage A as the major genotype based on the partial amplification and sequencing of gdh gene. In addition, Bahramdoust et al, [35] designed and evaluated a realtime PCR coupled with high resolution melting curve analysis (HRM) to detect and characterize G. duodenalis in humans and dogs, and reported assemblage A as the major genotype in humans. Although assemblage A seems to be the predominant genotype in humans in Iran [37][38][39], there are controversial results, as well.…”

Section: Plos Onementioning

confidence: 99%

Multigene typing of Giardia Duodenalis isolated from tuberculosis and non-tuberculosis subjects

et al. 2023

Self Cite

View full text Add to dashboard Cite

Giardia duodenalis is a cryptic protozoan, which has eight assemblages (A-H). Assemblages A and B are the main genotypes reported from humans with probable anthroponotic and zoonotic transmission. The current study aimed to characterize G. duodenalis assemblages in tuberculosis (TB) patients and healthy subjects using multilocus genotyping (MLG). Thirty Giardia-positive stool samples, which were obtained from TB patients and healthy subjects were included in the study. After total DNA extraction, three β-giardin (bg), triosephosphate isomerase (tpi), glutamate dehydrogenase (gdh) genes were amplified and sequenced. Obtained sequences were compared to the GenBank database to characterize assemblages. Phylogenetic analysis using Maximum Likelihood (ML) and Tamura 3-parameter was performed for each gene. From 30 Giardia-positive subjects, 17 (57%) and 13 (43%) were from healthy and TB-infected subjects, respectively. There was no significant co-existence of Giardia and tuberculosis (P-value = 0.051). In addition, 14 (46.7%) and 16 (53.3%) of Giardia isolates were from asymptomatic and symptomatic subjects, respectively. PCR amplification was successful in 25 single samples (83.3%) consisted of 20 for tpi, 15 for bg, and 13 for gdh genes. Accordingly, 13/25 (52%) and 8/25 (32%) belonged to assemblage A and assemblages B, respectively, whereas 4/25 (16%) were either assemblage A or B with different genes at the same time. Significant correlation between assemblages and TB, age, and symptoms was not seen. The phylogenetic analyses represented no separation based on TB and gastrointestinal symptoms. Assemblage A was the predominant genotype in samples. The high frequency of assemblage AII indicated importance of anthroponotic transmission of Giardia in both healthy and TB patients. In addition, considering the exclusive reports of sub-assemblage AIII in wild ruminants, the presence of AIII in the current study have to be carefully interpreted. The inconsistency between the assemblage results of either bg or gdh loci with tpi gene signifies the insufficiency of single gene analysis and the necessity for MLG in molecular epidemiology of G. duodenalis.

show abstract

“…Application of HRM assay has been reported for DNA sequence variations and genotyping of protozoan [31] and helminth [17] parasites. In this method, a close-tube allows detection of any alteration in DNA with simple screening, but it requires a small amplicon, which makes the selection of primers less exible [32].…”

Section: Mean Of T M and Melting Curve Analysis Of S Stercoralis Isol...mentioning

confidence: 99%

Application of Nested-qPCR-High Resolution Melting (HRM) Technology on Strongyloides stercoralis Isolates from Iran

Fakhrieh-Kashan,

Fotouhi-Ardakani,

Zahabiun

et al. 2024

Acta Parasit.

View full text Add to dashboard Cite

PurposeStrongyloides stercoralis is a parasite with special characteristics presenting it as a unique nematode. Iran is an endemic area for S. stercoralis. In this study, nested-qPCR-High Resolution Melting (HRM) technologies were applied on some human isolates of S. stercoralis from this country by focusing on evolutionary genetics analysis. MethodsTwelve human isolates of S. stercoralis were collected from four endemic provinces of Iran. Genomic DNA was extracted from a single lariform larva for every isolate. Using speci c primers targeting partial regions in cox1 gene, nested-qPCR-HRM was performed and melting-curve pro les were analyzed alongside with evaluation of genetic proximity and phylogenetic analysis using MEGA7 and DnaSP5 software. ResultsMelting temperature (T m ) values of the isolates were 77.9 ˚C -78.3 ˚C. All isolates from Guilan, Mazandaran, and Khouzestan Provinces shared T m values of 78.2 ˚C to 78.3 ˚C, while the isolates from Hormozgan Province showed T m values of77.9 ˚C, 78.0 ˚C, and 78.1 ˚C. The phylogenetic tree illustrated that the sequences of the current study included nine haplotypes. Tajima's D index analyses showed that cox1 gene in S. stercoralis isolates was negative (Tajima's D = − 0.27). ConclusionThe isolates were divided into ve temperature groups. Although compare to PCR sequencing HRM assay identi ed more limited genetic changes, but it revealed that the mean of T m of the isolates from Hormozgan Province was lower than those of other provinces, and represented speci c haplotypes for this geographical region on the phylogenetic tree.

show abstract

Development of HRM real-time PCR for assemblage characterization of Giardia lamblia

Cited by 10 publications

References 52 publications

Prevalence of Giardia duodenalis in dogs and cats: Age-related predisposition, symptomatic, and asymptomatic cyst shedding

Prevalence of Giardia duodenalis in dogs and cats: Age-related predisposition, symptomatic, and asymptomatic cyst shedding

Multigene typing of Giardia Duodenalis isolated from tuberculosis and non-tuberculosis subjects

Application of Nested-qPCR-High Resolution Melting (HRM) Technology on Strongyloides stercoralis Isolates from Iran

Contact Info

Product

Resources

About