جستجوی مقالات مرتبط با کلیدواژه « genotyping » در نشریات گروه « دامپزشکی »

Giardia duodenalis is a zoonotic protozoan infecting various vertebrates such as humans and domestic animals. The aim of this study was to determine the frequency and genotypes of G. duodenalis using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in dogs of Urmia, Iran. Overall, 246 stool specimens were collected from 100 pet, 49 stray, and 97 shelter dogs in the Urmia, Iran. Totally, seven samples (2.48%) were microscopically positive in terms of Giardia cyst. The PCR-RFLP analysis revealed that three (1.21%) and two (0.83%) samples have the C and D genotypes, respectively. In addition, two samples (0.83%) were belonged to the AI sub-group. A significant association was determined between the frequency of Giardia infection and life style, age, and stool form of dogs. The findings of the study showed the high frequency of Giardia infection in stray dogs and the dogs under one-year-old. Furthermore, the C and D genotypes of G. duodenalis were predominant in dogs of Urmia, Iran.

Infertility of unknown etiology is considered a significant medical and health problem. This study focused on the role of the estrogen receptor alpha (ESRα) gene polymorphism, PvuII (rs2234693), and its effect on the amount of ESRα in the blood of women who cannot get pregnant for unknown reasons. A total of 184 females were evaluated, including 102 with unexplained infertility (UI) and 82 age-matched control females (with at least one living child and no history of infertility). Blood samples were collected, genomic DNA was isolated from blood samples, and the genotyping of the ESRα gene was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). ESRα expression levels were assessed by the ELISA. The study revealed that the mean serum level of ESRα was significantly higher in the case group than in the control group (P<0.05). Furthermore, the genotypes (TT, TC, and CC) and alleles (T and C) significantly influenced the plasma level of ESRα in the study population. Moreover, the presence of the C allele was considered a risk factor, and the polymorphism had a significant effect on ESRα expression level in women with UI.

Amebiasis is caused by Entamoeba histolytica, a protozoan that is found worldwide. The degree of pathogenesis of clinical isolates varies greatly. This study was aimed to molecular identification of E. histolytica in children using the nested polymerase chain reaction (nPCR), and then, a genotyping of positive E. histolytica isolates using the quantitative PCR (qPCR) assay through targeting serine-rich E. histolytica protein (SREHP) gene. For this purpose, a total of 50 bloody diarrheic stool samples were collected from the children attended to the Al-Zahraa’ Teaching Hospital and Alkut Hospital for Gynecology, Obstetric and Pediatrics (Alkut, Wasit, Iraq) were subjected to the present study from September to December 2021. Firstly, the extracted DNAs that amplified using specific primers through targeting 18S rRNA gene and tested using nPCR assay were revealed an overall 48% (24/50) positive samples for E. histolytica. For genotyping, our results were detected an existence of four different genotypes (I, II, III and IV) with a significant prevalence of Genotype-II (54.17%) when compared to Genotype-I (20.83%), Genotype-III (12.5%) and Genotype- IV (12.5%). In addition, results of melting temperature of targeted genotypes were 84ºC, 83 - 83.5ºC, 82.5ºC and 81ºC for Genotype-I, II, III and IV, respectively. In conclusion, molecular amplification of 18S rRNA gene was revealed the large prevalence of E. histolytica among bloody diarrheic children of study areas; while, amplification of SREHP gene was reflected the widespread phenotypic variation of the Genotype-II suggesting the high ability of this genotype to spread infection in children. In different endemic areas as Iraq, the utilization of high-resolution genotyping methods showed the extremely polymorphic genetic structure of this parasite.

Newcastle disease virus (NDV) is an avian pathogen that infects various birds worldwide. Recurrent outbreaks of ND consistently occurring in Iran cause substantial economic losses each year. The Northeast region of Iran has an extensive commercial poultry industry and is also a big exporter of poultry products to other countries. Therefore, consistent and dynamic surveillance of the NDV's prevalence in this geographic region is essential in controlling the disease.

The virulence of the virus is determined based on the sequence of Fusion (F) protein. However, though the Phosphoprotein (P) and Matrix (M) proteins of NDV are also involved in the evolution and pathogenicity of the virus, molecular evaluation of their genomic loci in the NDVs prevalent in Iran is limited. Here, we present data for the sequences of full-length P and M genes belonging to an NDV that caused the ND outbreak of 2011 in the Northeast of Iran.

The genomic sequences encoding full-length P and M proteins as well as that of F protein were amplified using PCR and sequenced by the Sanger sequencing. The obtained sequences, plus their translated proteins, were evalu-ated using various bioinformatics approaches, such as homology and phylogenetic analyses.

Phylogenetic analyses based on P, M, and F genes clustered our isolate together with VII.I.I GenBank se-quences from Iranian sources reported from 2011 to 2019, as well as with those reported from China. But our isolate showed less homology to vaccine strains commonly used in Iran.

Our study showed that, in addition to the newly evolving sub-genotypes, VII.1.1 variants are still circulating in the region. The weak homology in determinant regions between this strain and those used for vaccine pro-duction must be considered in vaccination programs. Further, the persistent presence of NDV genotypes already prevalent in the Far East in Iran highlights the importance of biosecurity management and dynamic surveillance in controlling ND.

Infectious bronchitis (IB), caused by infectious bronchitis virus (IBV), is one of the most important respiratory diseases in poultry. The implementation of preventive measures, including vaccination and biosecurity, is necessary for controlling the disease. To maintain biosecurity, it is important to identify the entry route of new viruses into a region and characterizing markers such as unique mutations that make viruses traceable. During a genotyping study for IBV infected commercial chicken flocks in Khorasan Razavi province, 11 viruses from 11 broiler and layer chicken flocks were detected in different cities by PCR. Sequencing of the S1 partial gene followed by phylogenetic analysis showed that eight viruses can be classified in GI-23 lineage (Is-Variant2), two viruses are classified in GI-1 lineage (Mass), and one virus is classified in GI-12 lineage (793B). Although detected viruses of GI-23 lineage are originated from Iran, seven viruses have synonymous (T954C and G1056A) and non-synonymous (C797T) mutations that have not been previously reported. It was found that the new genetic changes in Iranian IBVs of GI-23 lineage occurred in two different regions in Khorasan Razavi. In conclusion, this study indicates that the high prevalence of GI-23 lineage viruses in Iran may enhance the chance of virus mutations and the emergence of new viral strains, so effective vaccination and biosecurity measures are required to control the virus spread.

Chlamydia psittaci (C. psittaci) is an avian pathogen which its clinical symptoms of the disease may be varies from asymptomatic to several clinical symptoms, which include: conjunctivitis, an inflammation of the lining of the nose (rhinitis), sinusitis, diarrhea (dehydration), respiratory distress, yellow-green urine, loss of appetite, which may cause respiratory disorders in humans. Oropharyngeal(owner and birds) and cloacal (birds) swabs were taken from 54 captive psittacine birds and their owners who attended to veterinary clinics in Isfahan (Totally 108 samples).To study the prevalence of C. psittaci in captive birds and their owners using molecular detection assay (PCR),samples were collected during 2014 from a total of 10 various species of parrots. C. psittaci was identified in four species of birds (40%). Sequencing was performed to confirm the PCR positive results, demonstrating that all positive samples of C. psittaci belonged to genotype A, representing the first report of the presence of this genotype in Iran. The determination of this bacterium in captive psittacine birds presents that there is a potential risk for owners who live or have direct contact with them and that there is a feasibility of infecting other birds and humans.

The foot-and-mouth disease virus (FMDV) with a wide variety of genomes and complicated biology is one of the infectious agents that put the lives of animals at risk. Therefore, to introduce suitable strains for vaccine production, it is essential to constantly evaluate genetic changes of circulating viruses in field. Within 2014-2015, a total of 126 clinical specimens consisting of epithelial tissue and vesicular fluid from tongue, dental pad, and hoofs suspected of FMD virus were submitted to the Reference Laboratory for FMD in Razi Vaccine and Serum Research Institute, and 86 of them were identified as FMD virus type A using sandwich Enzyme-Linked Immunosorbent Assay (ELISA). This virus was isolated from 42 samples from 16 provinces using cell culture. Firstly, the coding region that produces the main part of viral capsid was amplified by Polymerase chain reaction (PCR). This part of the genome by 800 bp length was related to the 1D gene that synthesizes the VP1 protein. The phylogenetic analysis of VP1 coding region determined two distinct genotypes with more than 15% nucleotide differences. The first cluster consisted of closely related viruses registered in the GeneBank of neighboring countries, including Afghanistan, Pakistan, and Turkey. All samples in Cluster1 were determined as relative viruses with genotype Iran-05. In-vitro serological examination indicated an antigenic relationship between Cluster 1 viruses and routine vaccine strain (A-IRN-2013). The second cluster with only two members was genetically far from earlier ones and could be considered a separate genotype. Furthermore, it was revealed that cluster 2 has not been previously reported in Iran. Genetic tracing indicated that these viruses might have been originated from circulating viruses from India.  Antigenic evaluation exhibited that this group could not be cross-protected by the routine vaccinal strain (A-IRN-2013) used during the research period.

Over the last decade, diagnostic tools to detect and differentiate Fasciola species have improved, but our understanding of the distribution of haplotypes and population structure of this parasite is less clear. This study was designed to survey this gap in the F. gigantica epidemiology in Kermanshah province, western Iran from 2015 to 2017.Sixty-eight Fasciola isolates were collected from slaughterhouses from this province. We evaluated the PCR-RFLP assay of the ITS1 genes for the identification of Fasciola species using the RsaI enzyme. After Fasciola species identification, the partial sequence of mitochondrial NADH dehydrogenase subunit 1 (ND1) gene of F. gigantica was used for subsequent construction of the phylogenetic tree and network analysis.Based on the PCR-PRFLP profile, one (6.25%) of sheep isolates and 19 (39.60%) of cattle isolates were detected as F. gigantica, whereas 93.75% of sheep isolates, 60.40% of cattle isolates and all of the goat isolates were F. hepatica. In the 20 analyzed flukes, five ND1 haplotypes were detected. Statistically significant genetic differentiation was demonstrated between the Iran population and all the other populations. Evidence is presented for the existence of two well-separated populations: African and West Asian gigantica flukes and East Asian gigantica flukes.Genetic relationships among haplotypes were associated with geographical divisions. Also, our results have heightened our knowledge about the genetic diversity of F. gigantic, providing the first evidence for the existence of two well-separated populations of this parasite.

Avian infectious bronchitis (IB) is an infectious viral disease of chickens. The effective protection of chickens against many different infectious bronchitis virus (IBV) variants is not achieved unless the circulating genotypes in the region are identified and the cross-protection of the potential of vaccines in use is assessed.

In a monitoring program of IBVs, a new genotype was identified in the north of Iran, 2019. This work was conducted to isolate and characterize this new IBV genotype.

Tracheal tissues were collected from chickens showing signs of respiratory involvement. Specimens were homogenized and inoculated to the allantoic fluid of embryonated specific pathogen-free (SPF) eggs. Infectious bronchitis virus was detected using real time-polymerase chain reaction (RT-PCR). The hypervariable region of the IBV S1 gene was amplified for sequencing.

Positive samples were phylogenetically analyzed, and both positive isolates were clustered with Q1 IBV strains.

This is the first report of the Q1 outbreak in Iran. More investigations are needed to find the role of Q1 IBV in the respiratory disease complex of chickens.

Major histocompatibility complex (MHC) represents an important genetic marker for manipulation to improve the health and productivity of cattle. It is closely associated with numerous disease susceptibilities and immune responses. Bovine MHC, also called bovine leukocyte antigen (BoLA), is considered as a suitable marker for genetic diversity studies. In cattle, most of the polymorphisms are located in exon 2 of BoLA-DRB3, which encodes the peptide-binding cleft. In this study, the polymorphism of the BoLA-DRB3.2 gene in Holstein's calves was studied using high resolution melting curve analysis (HRM). Observed HRM results were compared to PCR-RFLP and direct sequencing techniques. Eight different HRM and seven different RFLP profiles were identified among the population studied. By comparing to sequencing data, HRM could completely discriminate all genotypes (8 profiles), while the RFLP failed to distinguish between the genotypes *1101/*1001 and *1104/*1501. According to the results, the HRM analysis method gave more accurate results than RFLP by differentiating between the BoLA-DRB3.2 genotypes. Due to the Co-dominant nature of the MHC alleles, HRM technique could be used for investigating the polymorphisms of genotypes and their associations with immune responses.

Avian infectious bronchitis (IB) is a highly contagious viral disease which affects the poultry industry. The virus exists in a wide variety of genotypes, and phylogenetic analysis has been used to classify infectious bronchitis virus (IBV) strains. The object of the study is a molecular characterization of circulating IBV in Afghanistan as a first study. The tracheal tissue specimens from 100 different commercial broiler flocks with respiratory distress in Afghanistan were collected during 2016-2017. After real-time reverse transcriptase-polymerase chain reaction (RRT-PCR), IBV-positive samples were further characterized. A 390 bp hypervariable spike glycoprotein gene segment was amplified using Nested PCR, sequenced, and analyzed. The results of real-time RT-PCR showed that 45/100 of the mentioned flocks were IBV positive. Phylogenetic analysis of all positive samples revealed that IBV strains were clustered into two distinct genotypes: LX4 (GI-19) (9/45) and IS-1494 like (GI-23) (34/45). Also, 2 of the 45 samples remained uncharacterized. It is the first study focusing on the molecular epidemiology of IBV in Afghanistan, extending our understanding of IB in the region. These results showed the high rate of IB infection in Afghanistan broiler farms and confirm the continuing monitoring of IBVs to modify the vaccination program.

Clostridium perfringens has been known as a cause of diarrhoea in dogs. The aim of this research was isolation C. perfringens by culturing and toxinotyping by PCR molecular method. In this research 151 dogs’ faecal samples were collected from northwest of Iran, 131 of which were apparently healthy, and 20 of which were diarrheic. These faecal samples were cultured on 5% sheep blood agar; the suspected colonies with double homolysis that using multiplex and single polymerase chain reaction (PCR) assay were admitted to detect toxinotypes of the isolates by specific primers. C. perfringens strains were isolated from 5/20 (25%) the diarrheic group and 31/131(23.8%) the non-diarrheic group. All isolates (36/151) were classified as C. perfringens type A (cpa). Fourteen isolates (38.8%) with cpatpeL- profile and one isolate (2.8%) had cpatpeL- toxin’s profile. More studies are needed to elucidate the epidemiology of C. perfringens in dogs and its role as a zoonotic agent and public health hazard. Based on author’s knowledge, this is the first study was performed in order to isolation C. perfringens and genotyping from dogs in Iran. The cpa gene was reported from one C. perfringens isolated from healthy dogs.