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Showing 26 results for Pcr
Parviz Parvizi , Elnaz Alaeenovin ,
Volume 11, Issue 2 (6-2011)
Abstract
Background & Objectives: Leishmania infantum is the causative agent of visceral leishmaniasis (VL). Based on isoenzyme typing of a few isolates from patients and domestic dogs, this parasite was considered to predominate in the Kaleybar focus of VL in northwest Iran. There is no report of sandfly infections in this region so this study was aimed to investigate the infection of the sandflies in the field. Methods: Sandflies were sampled using sticky paper and CDC traps. Morphological identifications were carried out based on characters of the head and abdominal terminalia. DNA extracted from sandflies abdomens and thoraxes. ITS-rDNA gene of parasite was detected and identified as Leishmania after sequencing. Results: Out of 146 sandflies 9 were found to be infected with Leishmania. For first time, three Leishmania species (L. infantum, L. tropica and L. major) were identified in sandflies simultaneously in the region. Among the all sandflies only one Phlebotomus perfiliewi (vector of VL) was found to be infected with L. infantum. All Isolates were confirmed by sequencing of ITS-rDNA gene. Conclusion: However, Leishmania tropica and L. major were found more than L. infantum in sandflies in Kaleybar but it could not conclude that these two species of Leishmania are causative agents of VL. Because many criteria should be considered to incriminate an agent or vector of the disease.
Parisa Tahmasebi, Fatemeh Maryam Sheikolslami , Parisa Farnia , Majid Sadeghizadeh, Rashid Ramazanzadeh, Mahdi Kazempoor, Mohammadreza Masjedi , Ali Akbar Velayati,
Volume 11, Issue 4 (12-2011)
Abstract
Background & objectives : Amikacin is one of the key second-line drugs for treatment of tuberculosis. Mutations at the codons 1400, 1401and1483 of the 16srRNA gene are associated with resistance to amikacin. The purpose of this study was to detect these mutations using PCR-RFLP method in multi-drug resistant (MDR) strains of Mycobacterium tuberculosis showing resistance to amikacin. Methods : Susceptibility of strains (n=100) against first and second–line anti-tuberculosis drugs was performed by proportional method. Based on antimicrobial resistance pattern 97 strains were analyzed by PCR-RFLP method. rrs1096 and rrs1539 primers were used to amplify a 460bp region of the rrs gene. Then, the PCR products were digested using Tai 1 and Dde1 restriction enzymes. The results were analyzed by the SPSS software using Chi-square test. Results : Based on results from proportional method, 63 strains (64.9%) were MDR (Multiple Drug Resistant), 26 (26.8%) and 8 (8.2%) strains were susceptible and non-MDR, respectively. Also, 13.4% and 6.1% of the strains were XDR (Extensively Drug Resistant) and TDR ( Totally drug resistant ) respectively. Using PCR-RFLP method, 7 (7.2%) strains were resistant and 90 (92.7%) strains were susceptible to amikacin respectively. Moreover, we found that the mutation at the codon 1400 was the most frequent mutations responsible for resistance to amikacin. Conclusion : The PCR-RFLP method can be used as a supplemental method to detect resistance to amikacin however to increase our knowledge, mutatuions in several number of codons in rrs gene need to be studied.
Zahra Derakhshani Nezhad, Fatemeh Maryam Sheikolslami, Parisa Farnia, Zahra Deilami Khiabani, Rashid Ramazanzadeh, Mahdi Kazempoor, Mohammad Reza Masjedi , Ali Akbar Velayati,
Volume 12, Issue 3 (9-2012)
Abstract
Background & Objectives: Ethambutol is one of the four main drugs in treatment of tuberculosis. The most common mutation associated with this drug resistance usually occurs in codon 306 of embB. The aim of this study was to detect ethambutol resistance using Allele-Specific PCR and Spoligotyping in various subtypes of Mycobacterium tuberculosis. Methods : 140 sputum specimens were collected from suspected TB patients. They were digested and decontaminated using Pettrof method before culturing them on LJ medium. Drug susceptibility testing was performed on 106 culture positive specimens using proportional method. DNA was extracted from the isolated organisms and subsequently subjected to Allele-Specific PCR to detect any mutationin embB306. Spoligotyping was then used to determine the subtypes. Results: Out of 106 cultures positive samples, 36 samples (33.9%) showed resistance to ethambutol using proportional method. Allele-Specific PCR assay identified 93 as sensitive and 13 (27.6%) as resistant strains. The results of PCR were in agreement with result of proportional method. The PCR method revealed that 61.5% of mutation occurred in the first and 38.5% in third nucleotides. Spoligotyping differentiated Mycobacterium tuberculosis strains into Beijing (10 9.4%), Bovis (2 1.8%), CAS (24 22.6%), EAI (1 0.9%), Haarlem (27 25.4%), LAM (5 4.7%), Manu (5 4.7%), T (27 25.4%) and U( 2 1,8%) families. The high frequency of mutation in embB gene was belonged to Haarlem, CAS and T subfamilies. Conclusion: Based on results current study, mutations in the genes other than embB might have occurred in the resistant strains that gave negative result in Allele-Specific PCR assay. Therefore other mechanisms of resistance to this antibiotic should be investigated.
Gholamreza Irajian , Reza Mirnejad, Mohammad Reza Jalali Nodoshan, Nafiseh Ghorbanpour,
Volume 13, Issue 1 (4-2013)
Abstract
Background & Objectives: Prostatitis is relatively one of the common diseases in elderly men. Treatment of this disease is difficult and because of frequent relapses, it provides complicated problems for patients and the physicians. Detection of reservoirs and determination of prevalence of involved microbes in prostatitis are important in the epidemiology and control of the disease. This study was conducted to determine the prevalence of Mycoplasma genitaliumin patients with prostatitis by sequencing and PCR-RFLP techniques. Methods : In this cross-sectional study, 200 paraffin-embedded prostate samples from patients with prostatitis during 2008-2011 were checked for M. gentialium. After cutting the tissues and homogenization, the genomic DNA was extracted and used as template in PCR. Primers targeting a 465 bp regions of 16S rRNA of M. genitalium were used in the assay. PCR products were sequenced and Cac8I, Bbs I, EcoRI, AluI, TaqI endonucleases were used in RFLP analysis. Results : Of 200 samples, 4 were positive in PCR. The results of DNA sequencing and RFLP confirmed the amplified genes corresponded to M. genitaliumG37. Conclusion : The Mycoplasma genome present in tissue samples of prostate showed this bacterium could be one of the risk factors for prostatitis in men. However large studies and control groups are needed to prove this finding.
Narghes Roozafzay, Leila Kokabee, Syrous Zeinali, Morteza Karimipoor,
Volume 13, Issue 1 (4-2013)
Abstract
Background & Objectives: Hemophilia A is an X-linked recessive bleeding disorder , which is caused by several different abnormalities in Factor 8 gene . Intron 22 inversion is the main causative mutation in 45-50% of severe hemophilia A patients. Moreover intron 1 inversion is responsible for >5% of severe hemophilia A cases . The goal of this study was to precisely analyse intron 1 inversion of Factor 8 gene in severe Hemophilia A patients who were negative for intron 22 inversion by Inverse Shifting-PCR (IS-PCR) method. Methods: In this experimental study, severe hemophilia A patients with less than 1% of normal FVIII activity level referred from Isfahan Seyedolshohada hospital were analyzed. After obtaining the consent from patients, genomic DNA from peripheral blood leukocytes was extracted. Then, Inverse-Shifting PCR method was exploited for detection of inversion of intron 1 of Factor 8 gene in patients who were negative for inversion intron 22 . Results : In 18 out of 32 patients who were negative for inversion intron 22, 2 (6.2%) had intron 1 inversion . Conclusion : The allele frequency of inversion of intron 1 at Factor 8 gene is in agreement with related studies. IS-PCR is a rapid, robust and cost effective method that can improve the molecular detection of inversion and is useful for analysis of hemophilia A patients, carrier testing and prenatal diagnosis.
Ali Pezeshki, Mostafa Rezaeian , Mitra Zarebavani,
Volume 13, Issue 2 (7-2013)
Abstract
Background & Objectives: Giardia duodenalis is the most common intestinal parasite with cosmopolitan distribution. This parasite has been found in the intestine of humans and other mammalian hosts including cats, dogs, cattle, sheep, deer, pigs and muskrats. It is postulated that animals maybe reservoir for human infection and viceversa. In present study, the possible genetic similarity between cat and humans Giardia and its probable zoonosis were investigated. Methods: Direct examination and formalin-ether concentration techniques were performed on stray and semi stray cat fecal specimens. Gradient sucrose method was applied for collection and purification of cysts and DNA extraction was performed by phenol-chloroform and CTAB (Cetyl trimethyl ammonium bromid ( methods. DNA of cysts could hardly be extracted after repeated freezing and thawing. Polymerase chain reaction (PCR) was performed for DNA amplification. In this study triosephosphate isomerase (tpi) gene was selected as a molecular marker. Two sets of primers (PM 290 and PM 924) were considered. Two restriction enzymes RsaI and AvaI were also used to determine restriction fragment length polymorphism (RFLP) for PCR fragments amplified by both primer sets. Results: Ten samples were positive for Giardia cysts which were examined for molecular investigation. Four cat isolates were amplified by PM 290. PCR-RFLP patterns were found to be similar to human isolates AC≠AF 069556 (subgroup of AC≠U 57897) with possibility of cross-transmission. Conclusion: Therefore the similarity of genomic characters of isolates of cat and human Giardia implies possibility of zoonosis and transmission of these protozoa from cat to human and vice versa.
Delsuz Rezaee , Gholamreza Zarrini , Mohammad Ahangarzadeh Rezaee,
Volume 14, Issue 1 (4-2014)
Abstract
Background & Objectives : Acinetobacter baumannii is an opportunistic Gram-negative pathogen with increasing relevance in a variety of hospital-acquired infections especially among intensive care unit patients. A. baumannii is mostly a cause of septicemia, pneumonia and urinary tract infection following hospitalization of patients. In this study antibiotic susceptibility pattern of A.baumannii isolates and molecular typing among isolates resistant by REP-PCR were determined. Methods : During study, the A. baumannii, were isolated from hospitals in Tehran. The isolates were identified using standard biochemical tests and antibiotic susceptibility was determined by the disk diffusion method. Extraction of DNA and molecular typing of isolates performed using CTAB method and REP-PCR, respectively. Results : In this study 75 A. baumannii isolates separated from patients with an average age of 51 ± 18.45 years . The highest resistance rate was against azteronam (97%), ceftazidim (93%), cefepime (93%), piperacillin-tazobactam (93%), ciprofloxacin (93%) and ticarcillin (93%) while the lowest resistance rate was against tigecycline (n= 51, 68%), followed by tobramycin (n=24, 32%), ampicillin-sulbactam (n=21, 28%), amikacin (n=16, 21%), and carbapenems (n=11, 15%). The REP-PCR in resistant of A. baumannii isolates showed that the genotypes of A, B and C are the predominant genotypes in the resistant antibiotics. Conclusion: This study showed a high percentage of resistance to antimicrobial agents among genotypes A, B, and C of the A. baumannii isolates therefore strategies to control the spread of A. baumannii isolates must be designed and evaluated.
Samira Sheikh Ghomi , Parisa Farnia , Mojtaba Darbouy ,
Volume 14, Issue 2 (7-2014)
Abstract
Background & objectives: The rapid identification of patients carrying resistant Mycobacterium tuberculosis (M.TB) isolates is important for effective tuberculosis therapy. Unfortunately, during the recent years considerable numbers of isolates showed resistant to Rifampin (RIF) and Isoniazid (INH). The aim of this study was to rapidly identify resistant MTB isolates using molecular method. For this reason, the comparison between real-time PCR based on Taqman and HRM AssayS in detection of rpoB, inhA and katG genes mutation in clinical isolates were performed and analyzed. Methods: The study carried out on Mycobacteriology Research Center (MRC) from 2012-2013. Classical susceptibility testing i.e., proportional method against INH and RIF was performed on eighty three M.TB isolates. Thereafter, multiplex and real-time PCR were performed on extracted DNA sample. The real-time PCR was based on Taqman and HRM assays. Mutation in genes rpoB, inhA and katG were detected. Results: In overall, based on proportional and multiplex PCR method, 47 and 35 isolates were resistant to RIF and INH, respectively. Thirty of strains were resistant to both RIF and INH. The agreement of real-time PCR using Taqman was 88% for resistant and 84% for susceptible isolates, whereas the agreement of HRM was 96% and 30%, respectively. The sensitivity and specificity of Taqman in comparison to multiplex were 84% and 88%, respectively. In addition, the sensitivity and specificity of HRM were 30% and 96%, respectively. Conclusion: Results documented that real-time PCR based on Taqman assay is more sensitive than HRM assay. Additionally, real-time PCR based on Taqman assay is a rapid, accurate and cost effective method in detection of Mycobacterium tuberculosis resistance.
Fatemeh Hadadi, Azar Sabokbar, Mahrouz Dezfulian ,
Volume 14, Issue 2 (7-2014)
Abstract
Background & objectives: Trichophyton rubrum is one of the most common pathogeniccause of dermatophytosis. One of the drugs which have been prescribed widely for fungal infections is terbinafine which belongs to allylamines group of antifungal agents. Recently molecular typing methods have been developed for answering the epidemiological questions and disease recurrence problems. Current study has been conducted on 22 isolates of Trichophyton rubrum obtained from patients randomly. Our aim was the investigation of correlation between genetic pattern and sensitivity to Terbinafine in clinical isolates of Trichophyton rubrum. Methods: Firstly the genus and species of isolated fungi from patients have been confirmed by macroscopic and microscopic methods, then, the resistance and sensitivity of isolates against drug have been determined using culture medium containing defined amount of drug. In next step fungal DNA has been extracted by RAPD-PCR (random amplified polymorphic DNA) with random sequences of 3 primers. Results: Each primer produced different amplified pattern, and using each 3 primers differences have been observed in genetic pattern of resistant and sensitive samples using each 3 primers, but there was no bond with 100% specificity. Conclusion: The 12 sensitive isolates which didn’t grow in 0.1 mg concentration of drug, also had limited growth at the low concentration of drug. Ten resistant isolates which grew in 0.1mg/ml of drug, in lower concentration of drug were resisted. RAPD analysis for molecular typing of Trichophyton rubrum seems to be completely suitable.
F Hadadi, A Sabokbar, M Dezfulian , A Bakhtiari ,
Volume 15, Issue 2 (7-2015)
Abstract
Background & objectives: Trichophyton rubrum is one of the most common pathogenic causes of dermatophytosis. One of the drugs prescribed for fungal infections is fluconazole which belongs to Azoles group of antifungal agents. Recently molecular typing methods have been developed for answering the epidemiological questions and disease recurrence problems. Current study has been conducted on 22 isolates of Trichophyton rubrum obtained from patients randomly. Our aim was the investigation of correlation between genetic pattern and sensitivity to Fluconazole in clinical isolates of Trichophyton rubrum .
Methods: Firstly the genus and species of isolated fungi from patients have been confirmed by macroscopic and microscopic methods. Then, the resistance and sensitivity of isolates against drug have been determined using culture medium containing defined amount of drug. In next step fungal DNA has been extracted by RAPD-PCR (random amplified polymorphic DNA) with random sequences of 3 primers.
Results: Each primer produced different amplified pattern, and differences have been observed in genetic pattern of resistant and sensitive samples using each 3 primers, but there was no bond with 100% specificity.
Conclusion: The 12 sensitive isolates which didn’t grow in 50µg/ml concentration of drug, also had limited growth at the lower concentration of drug. Ten resistant isolates which grew in 50µg/ml of drug, also showed resistant to lower concentration of drug. There are differences in genetic pattern of resistant and sensitive samples. RAPD analysis for molecular typing of Trichophyton rubrum seems to be completely suitable.
B Davoodi, Kh Onsory , M Heydari Nasrabadi,
Volume 15, Issue 2 (7-2015)
Abstract
Background & objectives : Ovarian cancer is the most common female reproductive cancer which is caused due to the malignant transformation of ovarian cells. This type of cancer is the fifth most common cancer among women and the primary cause of cancer deaths in the world. Axin2 gene is a tumor suppressor gene of the Axin family in WNT cycle which is essential for embryonic development. WNT proteins in this pathway have important intermediary role in cell messaging and in primary and secondary development of the embryo. Axin2 gene is activated as a negative feedback to prevent excessive proliferation of cells with simultaneous activation of WNT messaging. The aim of this study was to find the frequency of mutation in rs1133683 region of exon 5 in Axin2 gene and its relation with the risk of ovarian cancer.
Methods : In this case-control study, 100 patients with ovarian cancer together with equal number of same age as controls were collected from Imam Khomeini Hospital. DNAs were extracted from blood and tissue and then were investigated by PCR-RFLP. Data analysis was performed using software SPSS (version 19) using logistic regression.
Results : The results of study of mutation in rs1133683 region of exon 5 in Axin2 gene between two groups of case and controls indicated that there is no significant association between CT genotype with ovarian cancer (OR=1.26, 95%CI 0.70-2.27,p=0.43). Also no association was observed between TT genotype of Axin2 gene and ovarian cancer risk (OR=1.56, 95%CI 0.49-4.96, p=0.44).
Conclusion : Study of mutation in rs1133683 region showed that there was no association between TT genotype carriers of Axin2 gene and the risk of ovarian cancer.
Esmaeil Babaei, Vahid Montazeri ,
Volume 16, Issue 3 (10-2016)
Abstract
Background & objectives: According to the new theory of cancer stem cells, interruption in the self-renewal pathway of tissue stem cells can cause cancerous tumors. Current work has evaluated the role of self-renewal Oct-4 gene in thyroid tumors.
Methods: In this case-control study, the expression of Oct-4 gene has comparatively assessed between cancerous specimens, marginal tissues of tumors and non-tumoral nodules of thyroid using RT-PCR technique.
Results: Statistical analysis of data by one-way ANOVA showed that Oct-4 gene is significantly expressed in thyroid papillary carcinomas in comparison with tumor margin and non-tumoral nodules (p<0.05).
Conclusion: In conclusion, the dominant expression of Oct-4 gene in thyroid tumoral cells not only demonstrates the cancer stem cell theory but also shows its role in thyroid cancer appearance that can be used in differentiating thyroid papillary carcinomas from non-tumoral nodules as well as demarcation of tumors.
Mortaza Nourmohammadi, Hosein Hamidinejat, Mohammadreza Tabandeh, Saad Goraninejad, Somaye Bahrami,
Volume 17, Issue 3 (10-2017)
Abstract
Background & objectives: Toxoplasma gondii is a zoonotic obligate intracellular protozoan parasite that infects all warm-blooded animals as well as human worldwide. Determining the parasite genotype in intermediate hosts is crucial in evaluating the role of these types in human infections as wll as in prevention programs. Therefore, this study aimed to identify and detect the genotypes of Toxoplasma gondii in aborted fetuses of ewes in Lorestan province.
Methods: Identification of the parasite was performed on the brain and liver tissues of 142 aborted fetuses using a conventional PCR based on amplification of highly repetitive 529 bp region of the parasite genome. Genotyping of positive samples, which were isolated from the brain and liver, was performed by PCR-RFLP based on SAG2, SAG3 and GRA6 molecular markers.
Results: From a total of 142 samples obtained from brain and fetus, 10 cases (7%) were determined as positive samples based on conventional PCR. The precence of parasite DNA was also confirmed in the liver of 3 positive samples. Evaluation of RFLP pattern of amplified SAG2, SAG3 and GRA6 genes showed the presence of various types of parasites, incuding type I in 3 samples, type II in 2 samples and atypical type in 5 samples.
Conclusion: Isolation of types I, II and atypical type of T. gondii from ewes in Lorestan province suggests the need for greater attention to parasite transmission from livestock to human, particularly in pregnant women and people with weakened immune system.
Maryam Tajoadini, Babak Kheyrkhah, Kuomars Amini,
Volume 18, Issue 1 (4-2018)
Abstract
Background & objectives: Shigella species are one of the most common causes of dysentery and sometimes death, especially in children and those with immunodeficiency. The variety of causative agents (Shigella species) and the development of drug-resistant strains make it difficult to select an appropriate antibiotic for the treatment of shigellosis. One of the most important factors involved in the resistance of Shigella isolates is the presence of extended spectrum beta lactamases (ESBLs) genes. The aim of this study was to determine the frequency of blaPER, blaGES and blaVEB genes in Shigella sonnei isolated from patients with dysentery using multiplex-PCR method and to determine the antibiotic susceptibility patterns of these isolates.
Methods: A total of 60 isolates of Shigella sonnei were collected from different hospitals and medical diagnostic laboratories in Kerman province. Specimens from different age groups were cultivated in special media and confirmed by biochemical tests. The presence of blaPER, blaGES and blaVEB genes were investigated using specific primers and multiplex-PCR method. Antibiotic susceptibility test was performed by disc diffusion method based on CLSI standards.
Results: Multiplex-PCR results showed three samples had blaPER gene, but none of them had blaVEB or blaGES genes. Also, the results of antibiotic susceptibility test showed the highest resistance for amoxicillin- clavulanic acid (53.3%) antibiotic and the highest sensitivity for tetracycline (85%) antibiotic.
Conclusions: The results of the experiments indicated the presence of blaPER gene in Shigella sonnei isolates. In addition, the results showed high resistance of isolates to amoxicillin clavulanic acid and ceftriaxone antibiotics. Therefore, careful medical care and proper and timely use of appropriate antibiotics are essential to prevent the spread of resistant isolates.
Arezo Kasavandi, Maryam Bikhof Torbati, Kumarss Amini,
Volume 18, Issue 3 (10-2018)
Abstract
Background & objectives: Staphylococci are considered as one of the most important etiological agents of omphalitis. Due to the importance of early diagnosis of omphalitis in newborns, this infection can be diagnosed by novel techniques such as multiplex PCR which is rapid, cost- effective and more accurate than microbial culture. The aim of this study was to determine the frequency of Staphylococcus aureus, S. epidermidis and S.hominis species in umbilical cord infection in newborns.
Methods: In the present study, 45 umbilical cord samples were collected from Shahid Afzali pour hospital in Kerman, Iran. Followed by DNA extraction, Multiplex PCR reactions were performed using specific 16srDNA primers for S. aureus, S. epidermidis and S.hominis. Finally, PCR products were analyzed using electrophoresis and sequencing. Also, microbiological and biochemical differentiation tests were performed for the diagnosis of Staphylococci on all specimens.
Results: Amplification of 16srRNA genes for S. aureus, S. epidermidis, and S. hominis using Multiplex PCR demonstrated that the frequency of S. epidermidis ,S. aureus and S.hominis were 4.4%, 6.6% and 2.2% in the studied samples, respectively. The prevalence of staphylococcal isolates using differential tests was shown to be 33.3%.
Conclusion: This study indicated that, Multiplex PCR is a proper method for simultaneous identification of S. aureus, S. epidermidis and S.hominis species. Also, Staphylococci can be considered as a significant cause of umbilical cord infection in newborns. However, further studies urgently are needed to confirm this finding.
Mandana Mansour Ghanaie, Sherin Tabrizian Namin , Ehsan Kazemnejad-Leili , Hanyeh Bashizadeh Fakhar , Mohammad Asgari Galebin ,
Volume 19, Issue 1 (4-2019)
Abstract
Background & objectives: Chlamydia trachomatis is a gram negative bacterium and chlamydia infection, as a curable infection, is one of the most common sexually transmitted diseases (STD). With regard to the essential role of chlamydia in infertility, the study of the prevalence of asymptomatic cases is precious. The aim of this study was to determine of the prevalence of chlamydia trachomatis in endocervical samples in infertile women with PCR method.
Methods: In this cross sectional descriptive-analytical study, a total of 135 women between 20-40 years old with chief complaint of infertility that referred to Alzahra-Rasht hospital and private clinics were randomly selected. The endocervical specimen was prepared using a sterile swab and was transferred to the laboratory in PBS for performing PCR. . The results of PCR and collected data from checklists were statistically analyzed using SPSS16.
Results: Chlamydia trachomatis was detected in 19.3% of infertile women. There were no statistically significant differences between PCR results and the patient's age, type of infertility, obstruction in salpingography, family history and duration of infertility.
Conclusion: The results of this study revealed that chlamydia infection has a high prevalence and in order to reduce the complications of this disease, screening tests can be used as a part of the country's health programs.
Atefe Sarafan Sadeghi , Najmeh Ansari, Farzad Khademi, Reza Mir Nejad , Behnam Zamanzad ,
Volume 19, Issue 1 (4-2019)
Abstract
Background & objectives: In recent years, Acinetobacter baumannii has been shown to be associated with several nosocomial infections, including pneumonia, bacteraemia, urinary tract infections, wound infection and meningitis. This organism can survive in the hospital environment and rapidly develops resistance to many antibiotics. The molecular genotyping can increase our knowledge about the spread of A. baumannii strains from one hospital to another and their drug resistance. Therefore, this study aimed to determine the prevalence of antibiotic resistance profile as well as phylogenetic relationships of A. baumannii strains in Shahrekord teaching hospitals.
Methods: In this study, antibacterial susceptibility patterns of A. baumannii strains isolated from different clinical specimens (urine, blood, sputum) to amikacin, ampicillin/sulbactam, aztreonam, cefepime, ceftazidime, ciprofloxacin, gentamycin, imipenem, meropenem, norfloxacin, ofloxacin, piperacillin/tazobactam, tobramycin were tested using disk diffusion )Kirby-Bauer( method. Finally, genotyping of A. baumannii strains was performed using REP-PCR method.
Results: During this study, 50 samples of patients were identified as A. baumannii) 71%(, and their drug resistance rates were assessed. All A. baumannii strains were resistant to ceftazidime and cefepime and also a high rate of resistance to aztreonam, norfloxacin, ciprofloxacin, amikacin, imipenem, gentamycin, and ampicillin-sulbactam were observed. On the other hand, our results demonstrated nine genotype groups among A. baumannii strains based on REP-PCR method.
Conclusion: Due to the high prevalence of antibiotic resistance among isolated A. baumannii strains, similarities between different genotypes and the dispersion of these genotypes in different parts of Shahrekord hospitals, the implementation of infection control programs in different parts of the hospital is necessary.
Khadijeh Khanaliha, Farah Bokharaei-Salim, Mohsen Sadeghi, Borna Salemi,
Volume 22, Issue 3 (10-2022)
Abstract
Background & objectives: Toxoplasma gondii is an obligate intracellular parasite with global distribution. Diagnosis of Toxoplasma gondii infection with high sensitivity and specificity is very important in managing and treating this disease. The purpose of this study is serological and molecular investigation of toxoplasmosis using B1 gene in HIV- positive patients referred to hospitals affiliated with Iran University of Medical Sciences.
Methods: In this study, 660 blood samples were collected from HIV/AIDS- positive patients referred to hospitals affiliated with Iran University of Medical Sciences. Patient samples were examined for the presence of IgG and IgM antibodies against Toxoplasma gondii using an ELISA kit. Genomic DNA was extracted from the patient's serum, as well as peripheral blood mononuclear cell (PBMC) and whole blood samples, and then Real time-PCR was performed.
Results: Although IgG antibody against Toxoplasma gondii was positive in 158 (23.9%) patients out of 660 HIV- positive patients, IgM antibody was positive in 5 (0.76%) patients. The results of Real-Time PCR showed that 7 (1.06%) patients were positive in PBMC samples, of which five patients were positive for IgM antibodies against Toxoplasma gondii while two patients had high- level Toxoplasma IgG antibody titers.
Conclusion: The results of the study indicate that the Real-time PCR method using PBMC DNA samples is a suitable method for the diagnosis of toxoplasmosis. This method, together with the antibody test, especially the high titer of Toxoplasma IgG antibodies, can be helpful in the diagnosis of toxoplasmosis.
Narges Chitsaz, Ahmad Reza Meamar, Elham Razmjou, Soheila Shafaghi-Sisi, Maryam Alipour, Maryam Sadeghi, Zahra Rampisheh, Zeinab Ghasemi, Rasoul Aliannejad, Alireza Badirzadeh,
Volume 23, Issue 4 (1-2024)
Abstract
Background: Pneumocystis jirovecii (P. jirovecii) causes Pneumocystis pneumonia (PCP) in people, especially the immunocompromised ones. It is also one of the serious causes of numerous lung problems in affected patients. Since documented data about P. jirovecii is not available in patients with pulmonary infections in Tehran, this study aimed to investigate the molecular epidemiology and parasitology of Pneumocystis to determine the frequency of the organism infection.
Methods: Bronchoalveolar lavage (BAL) samples were collected for 367 patients hospitalized in the lung department of Shariati Hospital in Tehran from July 2022 to July 2023. The samples were analyzed using Giemsa staining and molecular methods. After DNA extraction from samples, Nested polymerase chain reaction (Nested PCR) was employed for the amplification of the 18SrRNA gene and identification of P. jirovecii. The PCR products of Nested PCR were sequenced for final confirmation.
Results: Out of 367 samples, only one sample (0.27%) and 28 samples (6.7%) were found to be positive through parasitology and NestedPCR analysis, respectively. P. jirovecii was detected in seven (25%) and 21 (75%) immunocompromised and immunocompetent patients, respectively. Fever, shortness of breath and dry cough were the most common clinical symptoms among patients with Pneumocystosis. Patients with pulmonary disorders are prone to colonization by pneumocystis, which increases the risk of pneumocystosis and makes them a reservoir for transmission to susceptible people.
Conclusion: It can be concluded that patients with distinct lung disease are prone to colonization by Pneumocystis and, importantly, are at risk of infection. Also, according to the current study, Nested PCR was a suitable method for detecting P. jirovecii organisms because it had a very high sensitivity and specificity.
Morad Beiranvand, Hossein Hamidinejat, Somayeh Bahrami, Mohammad Reza Tabandeh, Meysam Makki,
Volume 24, Issue 2 (7-2024)
Abstract
Background: A zoonotic obligate intracellular protozoan parasite, Toxoplasma gondii, infects all warm-blooded animals as well as humans worldwide. Identification of the level of infection in intermediate hosts gives us an important data about understanding the role of this parasite in human health as well as estimating the economic loss in livestock. Therefore, the main aim of this study was the isolation and identification of T. gondii from aborted goat fetuses by PCR in Lorestan province.
Methods: From autumn 2023 to summer 2024, the brain and liver of 100 goat fetuses were examined for T. gondii by PCR based on the amplification of 529 base pair fragments from repetitive regions of the parasite genome. The study was performed in three aborted fetus groups, less than 2 months, 2 to 4 months and more than 4 months.
Results: From a total of 100 examined samples, conventional PCR detected the T. gondii infection in 6 (6%) and 2 of the brain and liver fetuses respectively.
Conclusion: This study shows a notable level of infection in goat fetuses, and as a result, T. gondii should be considered an important agent involved in the abortion of goats in the Lorestan province of Iran. On the other side, it is necessary to pay more attention to the risk of transmission of this parasite from farmed animals to humans, especially pregnant women and people with suppressed immune system.
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