جستجوی مقالات مرتبط با کلیدواژه « PCR-RFLP » در نشریات گروه « دامپزشکی »

Burkholderia mallei is the main cause of glanders as a dangerous contagious zoonosis disease that is mostly observed in single-hoofed animals, especially horses. Modern molecular techniques have been recently employed to improve epidemiology for identifying and searching for strains of this bacterium at different times and locations. Due to the unknown number of circulating strains and lack of preventive methods, glanders is still observed in the form of epidemics. The present study aimed to evaluate six field isolates plus two laboratory strains of Borkolderia mallei and Burkholderia pseudomallei using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. All the isolates and strains were microbially cultured in the glycerol nutrient and glycerol agar media. The individually grown colonies of the bacterium were used in the biochemical tests. The DNA of isolates was extracted by boiling, and the PCR-RFLP test was conducted on their genome. Finally, the bacterium was injected into guinea pigs to induce the Straus reaction. The biochemical assays (or bioassays) confirmed the isolates as Burkholderia mallei. The PCR-RFLP assay demonstrated a product for Burkholderia mallei with a length of 650 bp. Nevertheless, 250 and 400 bp were produced for Burkholderia pseudomallei. The swollen scrotum pointed to the occurrence of the Straus reaction. The PCR-RFLP is a proper differential diagnosis technique for B. mallei; moreover, it is a suitable method for differentiating between Burkholderia mallei and Burkholderia pseudomallei. This technique can detect Burkholderia mallei in a short time with high precision and sensitivity.

Identification of genotypic characteristics and pathogenicity of Staphylococcus aureus isolates is very important in the epidemiological study of its related diseases.

The present study was done to compare the S. aureus isolates from different sources on the basis of virulence gene properties, biofilm production ability, and phylogenetic variations.

Seventy S. aureus isolates (including 25 human, 25 raw milk, and 20 pet animal isolates) were subjected to slime production ability testing, polymerase chain reaction (PCR) detection of 14 different virulence genes, and DNA fingerprinting using restriction fragment length polymorphism (RFLP) of coa gene PCR products.

Among 70 S. aureus, 64 (91.4%) isolates were slime producers on Congo red agar (CRA) medium. The spa and icaD virulence genes were present in all isolates and the seD and etaA genes were not detected in any of the isolates. In total, 22 different virulence gene patterns and nine distinct clusters of coa-PCR-RFLP were identified among isolates.

According to the results, S. aureus strains of human origin showed a significant association with specific virulence gene profiles and genotypes. seB and seC were the most responsible genes for S. aureus enterotoxin among human and animal isolates, respectively. Coa-RFLP showed partially appropriate results in the classification and source detection of S. aureus isolates.

Fungal infections are currently causing health issues all over the world, among which are Candida species that cause cutaneous infection. Numerous dermatological studies concentrated on a single species. However, the virulence factors and the spread of specific candidiasis in specific areas have remained poorly understood. Therefore, the current study was designed to shed light on Candida tropicalis, which has been identified as the most prevalent yeast among Candida non-albicans species. Forty specimens were collected from patients with cutaneous fungal infection (25 females and 15 males) and underwent examination. According to conventional identification based on macroscopic and microscopic examinations, eight isolates were identified as C. tropicalis from Candida non-albicans. Molecular diagnosis for internal transcribed spacers (ITS1 and ITS4) using conventional polymerase chain reaction (PCR) yielded an amplicon of 520 bp for all isolates. Further investigation of PCR-restriction fragment length using Mitochondrial sorting protein; Msp1 enzyme revealed two bands of 340 and 180 bp. The ITS gene sequence in one isolated species was found to be 98% identical to C. tropicalis strain MYA-3404 chromosome R ATCC CP047875.1. Another isolate shared 98.02% identity with C. tropicalis strain MA6 18S ribosomal RNA gene DQ666188.1, indicating C. tropical species identity, implying that non-Candida species should be considered when diagnosing candidiasis. This study demonstrated the significance of Candida non-albicans, particularly C. tropicalis, in terms of pathogenic potential, the ability to cause potentially fatal systemic infections and candidiasis, and acquired flucozonal resistance with a high mortality rate.

Multiple sclerosis (MS) is a chronic autoimmune disorder of the central nervous system. Genetic and environmental factors have important roles in the induction, onset, and progression of MS. In this study, the IL-21 genotype (rs2055979) (G/T) in Iraqi MS patients was compared with a healthy control group to investigate the possible association of any particular genotype or allele with multiple sclerosis. This study included 70 patients with relapsing-remitting MS and 50 healthy individuals as control. Following the extraction of genomic DNA, polymerase chain reaction-restriction fragment length polymorphism, the frequencies of genotypes, and alleles were calculated and statistically analyzed. The results of the study revealed a significant reduction in the distribution of the wild homozygous genotype (GG) in MS patients, in comparison to a healthy control group (14.3% vs. 34 %; 0.0129 at P≤0.05; odds ratio [OR] 3.0909, 95% confidence interval [CI]: 1.2704-7.5203). However, MS cases and controls did not differ significantly in neither GT nor TT genotypes, 62.9% (OR 0.6402, 95% CI: 0.3064-1.3374) and 52% (OR 0.5494, 95% CI: 0.2074-1.4557), respectively. The data of allele frequencies in patients and controls showed that the G allele frequencies were 0.46 vs. 0.60 in patients and controls, respectively, while T allele frequencies were 0.54 vs. 0.40 in patients and controls, respectively. The current conclusions indicated that in the study group, the GG genotype of IL-21(rs2055979) could be related to MS.

Forty-eight isolates of hydatid cyst collected from sheep and camels slaughtered in different areas of Iran were analyzed by polymerase chain reaction and restriction fragment length polymorphism techniques. The results indicated that the isolates could be categorized into two distinct and uniform genotype grouping, the camel/dog strain,group (Genotype 6) and cosmopolitan common sheep strain group (Genotype 1) of E.granulosus. These isolates can potentially act as a risk factor for human health in the country.