Molecular Characterization of Trichophyton Verrucosum Strains İsolated From Cattle by PCR-RFLP

Abstract Background: Clostridioides difficile is the main cause of healthcare-associated diarrhea worldwide. It is proposed that certain C. difficile toxinotypes with distinct pathogenicity locus (PaLoc) variants are associated with disease severity and outcomes. Additionally, few studies have described the common C. difficile toxinotypes, and also little is known about the tcdC variants in Iranian isolates. We characterized the toxinotypes and the tcdC genotypes from a collection of Iranian clinical C. difficile tcdA+B+ isolates with known ribotypes (RTs).Methods: Fifty C. difficile isolates with known RTs and carrying the tcdA and tcdB toxin genes were analyzed. Toxinotyping was carried out based on a PCR-RFLP analysis of a 19.6 kb region encompassing the PaLoc. Genetic diversity of the tcdC gene was determined by the sequencing of the gene.Results: Of the 50 C. difficile isolates investigated, five distinct toxinotypes were recognized. Toxinotypes 0 (33/50, 66%) and V (11/50, 22%) were the most frequently found. C. difficile isolates of the toxinotype 0 mostly belonged to RT 001 (12/33, 36.4%), whereas toxinotype V consisted of RT 126 (9/11, 81.8%). The tcdC sequencing showed six variants (35/50, 70%); tcdC-sc3 (24%), tcdC-A (22%), tcdC-sc9 (18%), tcdC-B (2%), tcdC-sc14 (2%), and tcdC-sc15 (2%). The remaining isolates were wild-types (15/50, 30%) in the tcdC gene.Conclusions: The present study demonstrates that the majority of clinical tcdA+B+ isolates of C. difficile frequently harbor tcdC genetic variants. We also found that the RT 001/ toxinotype 0 and the RT 126/ toxinotype V are the most common types among Iranian isolates. Further studies are needed to investigate the putative association of various tcdC genotypes with CDI severity and its recurrence.

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Molecular Analysis of Aquaglyceroporin 1 Gene in Non-Healing Clinical Iso¬lates Obtained from Patients with Cutaneous Leishmaniasis from Central of Iran

Journal of Arthropod-Borne Diseases ◽

10.18502/jad.v13i2.1240 ◽

2019 ◽

Author(s):

Yasaman Alijani ◽

Saeedeh Sadat Hosseini ◽

Salman Ahmadian ◽

Sonia Boughattas ◽

Gilda Eslami ◽

...

Keyword(s):

Gene Expression ◽

Cutaneous Leishmaniasis ◽

Molecular Characterization ◽

Significance Level ◽

Detection And Identification ◽

Pcr Rflp ◽

Leishmania Spp ◽

Sensitive Isolate ◽

The One

Background: Regarding the antimonial-resistant of Leishmania spp., understanding of related mechanism is necessary. One of the most important involved molecules is aquaglyceropin1 (AQP1). The aim of this study was molecular analysis of AQP1 gene from antimonial-resistant clinical isolates and its expression. Methods: Overall, 150 patients with cutaneous leishmaniasis referring to the reference laboratories of Yazd and Varzaneh,, located 105km southeast of Isfahan and 240km away from Yazd, were assessed from Jun 2015 to Dec 2017. After sampling, staining was done and evaluated for Leishman by microscope. Samples were collected in RNAlater solution for gene expression analysis in non-healing isolates. DNA extraction was performed from each slide with Leishman body. All patients with L. major isolates detected by ITS1-PCR-RFLP were followed for finding the resistant isolates, consequence of molecular characterization of AQP1 using PCR-RFLP. Gene expression of AQP1 from all resistant isolates was assessed in comparison with the one in a sensitive isolate. Statistical analysis was done using SPSS. The significance level was considered ≤0.05. Results: Five isolates were detected as antimonial resistant. Molecular detection and identification were appeared that all were L. major. The molecular characterization of AQP1 showed G562A mutation. Gene expression of AQP1 in resistant isolates showed 1.67 fold higher than the sensitive isolate. Conclusion: We reported a new point mutation of G562A in AQP1 gene involved in molecular mechanism in resistant isolates.

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Molecular characterization of Ovar-DRB1 exon2 gene in Garole sheep resilient to gastrointestinal nematodes

Indian Journal of Animal Research ◽

10.18805/ijar.6710 ◽

2015 ◽

Author(s):

C. Paswan ◽

L. L.L. Prince ◽

R. Kumar ◽

C. P. Swarnkar ◽

D. Singh ◽

...

Keyword(s):

Disease Resistance ◽

Molecular Characterization ◽

Molecular Mechanisms ◽

Gastrointestinal Nematodes ◽

Exon 2 ◽

Pcr Rflp

Ovine MHC DRB1 exon 2 (Ovar-DRB1.2) gene is arguably one of the critical genes, responsible for disease resistance against parasite in animals. This study was carried out for indepth analysis of polymorphism in MHC DRB1.2 gene and to explore the underlying molecular mechanisms for the development of disease resistance in Garole. PCR-RFLP study revealed that the gene is polymorphic in nature. The frequency of allele ‘A’ for endonuclease SacI and Allele ‘B’ for endonuclease Hin1I were significantly higher in Garole population.

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Molecular characterization of 'Candidatus Phytoplasma solani' in celery: Case study in Futog

Ratarstvo i povrtarstvo ◽

10.5937/ratpov58-33227 ◽

2021 ◽

Vol 58 (2) ◽

pp. 66-71

Author(s):

Tatjana Popović ◽

Petar Mitrović ◽

Andrea Kosovac

Keyword(s):

Molecular Characterization ◽

Gene Detection ◽

Convolvulus Arvensis ◽

Visual Evaluation ◽

Leaf Yellowing ◽

Pcr Rflp ◽

Novi Sad ◽

Stolbur Phytoplasma

'Candidates Phytoplasma solani', known by its trivial name stolbur phytoplasma, is a plant pathogen infecting numerous crops in Serbia. Celery plants with prominent leaf yellowing and chlorosis, sporadically with tissue necrosis, were observed during August 2020 in Futog, situated in Novi Sad suburbia in Vojvodina. Total of 12 sampled celery plants, 8 symptomatic and 4 asymptomatic ones, were analysed for 'Ca. P. solani' presence. All symptomatic celery plants were infected with stolbur phytoplasma according to the stoll 1 gene detection and therefore were further subjected to multigene molecular characterization on three genes: tuf, stamp and vmpl. Combining molecular tools PCR/RFLP and sequencing reviled two 'Ca. P. solani' multilocus genotypes in celery: tuf-b/Rqg31/V14 and tuf-b/ Rpm35/V14, present in 4 samples each. Obtained results of the strain genotyping are in concordance with previous data on the 'Ca. P. solani' diversity on celery, but supplemented with genotyping of the vmpl gene. Outbreak of stolbur phytoplasma in the assessed locality in Futog is linked to tuf-b epidemiological cycle correlated in Serbia mainly with weed Convolvulus arvensis which was present in the subjected celery plot, and could have been the phytoplasma inoculum source. Visual evaluation of the symptom occurrence suggests on 10-15% of 'Ca. P. solani' affected celery plants scattered throughout the plot corresponding to the pathogen dispersal in crop by cixiid planthoper Hyalesthes obsoletus (Hemiptera: Auchenorrhyncha) associated with C. arvensis, main vector of stolbur phytoplasma in Serbia.

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