Showing 41 results for Pcr
M Sattari, Aa Imani Fooladi, Gh Godarzi,
Volume 3, Issue 1 (4-2009)
Abstract
Abstract Background and objectives: Pseudomonas aeruginosa as an opportunistic pathogen can establish lethal infections in immunocompromised patients or those exposed to predisposing factors. This bacterium contains a single polar flagellum causing motility, chemotaxis and colonization in acute phase of infection. The flagella filament is made up of a structural protein called flagellin. This study was aimed at determining The frequency of fliC gene in Clinical Samples. Material and Methods: In this study, a pair of specific primer for types of flagellin (a, b type) was designed and by using PCR method its structural gene (fliC) was recognized and amplified in clinical strains. Results: This original primer has appropriate efficiency in diagnostic of pseudomonas aeroginosa flagellum. Our study shows that 85% of the Clinical Samples have a fliC gene. Conclusion: This method can be applied to recognizing of the motile strains, and their antigenic typing, and complete amplification of fliC sequence in order to cloning and expression of recombinant flagellin. Key words: Pseudomonas aeruginosa, flagellin, fliC, PCR
S N Javid, E A Ghaemi, N Amirmozaffari, S Rafiee, A Moradi, T Dadgar,
Volume 3, Issue 1 (4-2009)
Abstract
Abstract
Background and objectives: With almost nine million new cases each
year, tuberculosis is still one of the most Life-threatening diseases in the
World. Distribution of drug resistant strains of M.tuberculosis has a lot of
importance. This research was carried out to determine the frequency of drug
resistance of M. tuberculosis in strains isolated in Golestan province.
Material and Methods: In this cross -sectional study, 104 isolate of
M.tuberculosis which isolated from patients referred to Gorgan tuberculosis
Health Center, in 2008 were studied. DNA was extracted by Boiling Method.
By using PCR method, we determine the M.tubeculosis strain and resistance
to Rifampin (Using IS6110 and Gene rpoB primers) and resistance to
Isoniazid (Using InhA and KatG primers). As a Gold Standandard,
“Proportional method” was performed for 45 Samples.
Results: 87 strains were identified as M.tuberculosis. 6.9% of them were
resistant to Isoniazid, 4.6% to Rifampin and 2.3% to both (MDR).Sensitivity
and Specifity of PCR method in detection of resistant to Isoniazid were
95.3% and 57.1% and for Rifampin were 94.7% and 33.3%.
Conclusion: We found that in our region, the MDR is not very common.
More than 16% of isolated strains from tuberculosis suspected patients were
MOTT, for this reason it is necessary to mention that use biochemical or PCR
method to determine M.tuberculosis is necessary.
Key words: Mycobacterium tuberculosis, MDR, PCR, Proportional method
, Golestan province.
H Davoodi, S R Hashemi, H F Seow, M Ghorbani,
Volume 3, Issue 2 (10-2009)
Abstract
Abstract Background and objectives: Paraffin-embedded tissues and clinical samples are a valuable resource for molecular genetic studies, but the extraction of high-quality genomic DNA from this tissues is still a problematic issue. In the Present study, the efficiency of two DNA extraction protocols, a commercial kit and a traditional method based on heating and K Proteinase was compared. Material and Methods: Fifty paraffin-embedded blocks of colon cancer tissues (more than 5 years old) were used to compare two methods of DNA extraction. DNA was extracted by traditional method using heat and commercial DNA extraction (Qiagen kit) method. Then the purity and concentration of extracted DNA were measured by Spectrophotometer. Two sequences of TLR4 “The most important receptors in innate immunity” were amplified by polymerase chain reaction. SH-1 ‘188bp’ and SH-2 ‘124bp’ were amplified and then the products were separated on a 2% agarose gel. Results: The results show that the yield of DNA by traditional method (297 mg/ml) is significantly (p<0.01) higher than Commercial kit (176mg/ ml). But traditional method has the lower OD ratio (1.2) Compared to Commercial method. The Amplification of the TLR4 gene sequences is more successful by the traditional method (p<0.01) compared with commercial method. The length of the sequence affects on the results of PCR in that short sequence is amplified more successful compared to the long sequence. Conclusion: The traditional method is more successful in PCR amplification and also simple and cheap. Therefore, we recommend using this method for DNA extraction taken from the paraffin-embedded blocks with more than 5 years old and selecting shorter sequence for better amplification in PCR. Key words: DNA Extraction, paraffin embedded tissue, PCR
S H Alizadeh Shargh, A Ghazanchaei, A A Ayetollahi, A Khandan Del, B Pourasghari, R Estakhri,
Volume 4, Issue 2 (10-2010)
Abstract
Abstract Bachground and objectives: Dientamoeba fragilis is a habitant protozoa in human colon causing clinical symptoms, such as local stomach pain, weight loss, lack of appetite and flatulence. It is important to diagnose this infection correctly and differentiate it from other Protozoa. In this study PCR and Iron Hematoxylin methods were used to detection of this protozoa in Chalous Medical centers refers in 2010. Material and Methods: The stool samples (n=302) of this cross-sectional study were selected via cluster random sampling. After wet mount study the samples were preserved in PVA (for staining) and Ethanol (for molecular). The samples were studied both Staining and Molecular methods. Sensitivity and specificity were assessed. Results: Of 302 samples, six of them are positive via staining method (1.99%) and five by molecular method. All negative results with staining method are also negative with PCR. Contamination with E.coli in 2 samples and with Balstocystis homonis were seen in one sample. Sensitivity and specificity of PCR was 85% and 100% respectively. Conclusion: The discrepancy between two methods maybe caused by observer errors in staining method and unsynchronized molecular and microscopic studies. Key words: Dientamoeba fragilis, PCR, Iron Hematoxylin, Chalous region
Biranvand E, Abedian Kenary S, Ghaheri A, Rezaee M S, Hasannia H, Nasrolahi M, Parsaee Mr, Ahanjan M, Biranvand B, Ahmadi Basiri E,
Volume 5, Issue 1 (4-2011)
Abstract
Abstract Background and objectives: Interferon-Gamma and interferon Gamma receptor (IFNγ ⁄ IFNγR1) are the main genes associated with susceptibility to tuberculosis. We aimed at studying on interferon-Gamma Gene polymorphism(- 56 C/T) in people suffered from tuberculosis (TB). Material and Methods: In this case-control study, the subjects were 62 individuals with TB and 74 healthy ones. Genomic DNA was extracted by DNA isolation kit(Roche Corporation), and genotype identification was performed by polymerase chain reaction-Restriction fragment length polymorphism (PCR-RFLP). Chi square and logistic regression, using SPSS soft ware (version 18), was used to compare genotype and alleles between case and control groups. Results: The frequency of TT genotype in tuberculosis patients and healthy person are 43.5% and 17.5%, respectively. Based on Logistic regression (odd ration 0.148, p=0.0006), there is significant difference between Case and Control. In addition, the frequency of T allele is, in case group, 62.09 % the difference between case and control is significant, based on Logistic regression (odd ratio: 0.418, P=0.028). Conclusion: It is implied that -56C/T is associated with IFNγR1 promoter in tuberculosis patient. It is found to be associated with increased susceptibility to tuberculosis. Key words: Tuberculosis, IFNγR, PCR-RFLP
Teyhoo M, Mobin H, Mozafari N A, Moadab S R, Sedigh Bayan Kh, Mones Rast Sh,
Volume 5, Issue 1 (4-2011)
Abstract
Abstract Background and objectives: Staphylococcus aureus is one of the most important etiological agents of hospital and community acquired infections. The enterotoxins and toxin shock syndrome toxin (TSST-1) are among the most common virulent determinants of this bacterium. They are also well-known for their super-antigenic properties. The incidence of TSST-1 producing strains is also very alarming. The aim of this investigation was to survey the prevalence of TSST-1 gene in the clinical isolates of S. aureus recovered from hospitalized patients in Shohada hospital of Tabriz, Iran. Material and Methods: During one year period, 1454 specimens obtained from hospitalized patients were investigated. After doing Isolation and purification, the isolates were identified by routine bacteriological methodologies.Their antibiotic susceptibility patterns were determined by agar disk diffusion method. Following genomic DNA extraction by boiling method, the presence of TSST-1 gene was analyzed by PCR. Results: A total 100 S. aureus isolates were recovered (6.87%). Antibiogram results indicate that all of the isolates are sensitive to linzolid 83% of them are resistant to meticillin. The prevalence rate of TSST-1 gene in the isolates is 20%. Conclusion: The high prevalence of TSST-1 gene in studied S. aureus strains and their circulation in the community can have a potentially alarming effect on general health of community. Key words: Staphylococcus aureus, TSST-1, Antibiotic resistance, PCR
Moradi Av, Azadfar S, Fatemehcheraghali, Javid N, Ghaemi A, Tabarraei A,
Volume 5, Issue 2 (10-2011)
Abstract
Abstract Background and objectives: Mumps virus is one of the first known causative agents of meningitis in children. On-time diagnosis is the first step in treating meningitis. We aimed to evaluate Mumps virus meningitis in children in Gorgan, Iran Material and Methods: CSF and blood samples were taken from children with meningitis, Jun 2008 till Sep 2010. For 40 samples with negative bacterial culture, Extraction of viral RNA was carried out and Real-time PCR was performed for detection of Mumps virus. Demographic, clinical, biochemical and cytological data were collected. We run SPSS version 18 to analyze the data, using Chi Square (p<0.05). Results: three (7.5 %) samples have Mumps virus, two boys and one girl. All three positive cases have 0.5-1 degrees Celsius fever and vomiting but no bulging fontanel. They have not Kernig, Rodor, Brudzinski’s sign, hepatosplenomegaly, lymphadenopathy, pharyngitis and rash. ESR is higher than normal in all positive cases and CRP is positive in two cases. Protein of CSF in one case is higher than normal range. Conclusion: meningitis is an emergency condition therefore, molecular diagnostic techniques are recommended for early diagnosis and intervention. Key words: meningitis, mumps virus, cerebrospinal fluid, Real-Time PCR
Mousazade Moghadam M, Babavalian H, Mirnejad R, Shakeri F,
Volume 6, Issue 1 (4-2012)
Abstract
Abstract
Background and objectives: Genomic DNA extraction of bacterial cells is of processes performed normally in most biological laboratories therefore, various methods have been offered, manually and kit, which may be time consuming and costly. In this paper, genomic DNA extraction of Staphylococcus aureus was investigated using some laundry detergent brands available in Iran to achieve a rapid and cost effective method.
Material and Methods: five-enzyme Taj brand, three-enzyme Saftlan brand ,and Darya and Pak brands without enzyme were used in the concentrations of 10, 20, 40, 80 mg/L. Afterwards, in order to evaluate the efficiency of extracted DNA in downstream processing, PCR test was performed for femA gene in the genome of Staphylococcus aureus.
Results: DNA extraction using different concentrations of the brands show that extracted DNA using 40 mg/L Saftlan and Taj brand powders have the best results according concentration (µg/ml) and purity (A260/A280) parameters. These parameters are 387.5 1.88 (Taj), 254.1 2.80 (Softlan), 396.6 1.95 (Manual) and 423.3 2.2 (Kit), respectively. Afterward, the PCR test results by show that DNA extraction using laundry detergents has no effect on its efficiency in order to be used in downstream processes.
Conclusion: These results indicate that the proper concentrations of laundry detergents can be used to extract genomic DNA with similar efficiency to kit and manual extraction methods.
Key words: Bacterial genome, DNA extraction, laundry powder, PCR, Staphylococcus aureus
H Samadikafil, M Mohebatimobarez, M Forouzandehmoghadam,
Volume 7, Issue 1 (4-2013)
Abstract
Abstract
Background and Objective: Enterococci are Gram-positive members of human gastrointestinal flora,in Dairy products and environment. they have emerged as important causes of opportunistic nosocomial infections in recent years. In this study we aimed to investigat and compare the efficiency of MALDI-TOFmass spectroscopy method through Biochemical and Molecular methods for detecting Enterococcus faecalis and Enterococcus faecium.
Materials and Methods:seventhy five clinical samples were collected for biochemical, molecular and mass spectroscopy investigations. Samples were treated with Esculin hydrolysis, Catalase, Pyrrolidonylaminopeptidase, 6.5% NaCl solution, motility, 0.04% Tellurite, L-Arabinose and Sorbitol. Using specific primesallele specific PCR was used.The samples were then analyzed by MALDI-TOF mass spectroscopy and Biotyper 3 software.
Results:Enterococcus faecium andEnterococcus faecaliswere detected in thirty and forty two samples, respectively whereas three samples showed both bacterial infections. Using biochemical analysis, two E.faecium isolates were Arabinose negative and one E. faecalis isolates was Telliurite negative. All sampleswere showed correct bands in PCR results but twoof them didn't show clear bands(on agarose gel). In mass spectroscopy analysis all strains were correctly detected and well defined.
Conclusion: According to our results, MALDI-TOF mass spectrometry in comparison with Molecular and Biochemical Methodscould be a reliable and accurate method that can easily and quickly identify and differentiate Enterococcus faecium and Enterococcus faecalisin clinical samples.
Key words:Enterococcus faecalis, Enterococcus faecium, MALDI-TOFmass spectrometry,PCR
M Fakhar, E Ahmad Pour,
Volume 7, Issue 1 (4-2013)
Abstract
Abstract
Visceral leishmaniasis (Kala-azar) is a systemic infection disease that can be diagnosed by some invasive procedures such as splenic, liver biopsy or bone marrow aspiration, whichare determined as the gold standards for diagnosing of this disease. At present, a variety of noninvasive tests having different specificities and sensitivities are available for the diagnosis of visceral leishmaniasis. Direct agglutination test (DAT) can be an appropriate and applicable method provided that proper antigens are prepared. The rapid rK39 strip test (for detection of antigen) can be used for diagnosis of visceral leishmaniasis (VL), which is suitable for acute forms of disease in the field. Other tests, such as rapid KATEX strip test (for detection of antigen) and polymerase chain reaction (PCR), which are recently recommended for diagnosis and prognosis of visceral leishmaniasis, are the simple, inexpensive and easily available under field conditions.This review article focuses on different, novel and current procedures for the diagnosis of visceral leishmaniasis.
Key words: Laboratory diagnosis,visceralleishmaniasis, Kala-azar,rk39, Katex, PCR
H Bashi Zadeh Fakhar, R Faraji, M Ghane, M Jafarpour, B Ashoorizadeh,
Volume 7, Issue 3 (10-2013)
Abstract
Abstract
Background and Objective: Much research has shown that Human Papiloma Virus (HPV) plays an important role in cervix cancer and it is the cause of 99% of cervix cancer worldwide. Lots of research has been done to find a proper method for HPV diagnosis and screening in patients with genital warts. This study aimed at comparing PCR method with Pap smear test in HPV screening.
Material and Methods: Considering the presence of DNA of HPV, 45 vaginal and cervix swap samples of women with genital warts were tested by means of specific PCR and Pap smear from September 2010 to April 2011.
Results: Out of 45 vaginal and cervix swap samples of women suffering genital warts, 37 samples (82.2%) are positive. Of 45 Pap smear samples, 13 (29%) are neoplasia and 32 (71%) normal.
Conclusion: The difference between the results of PCR and Pap smear is due to low specification and sensitivity of Pap smear. Thus it is recommended using diagnostic PCR method in addition to Pap smear in order to promote the quality of screening in individuals with genital warts.
Keywords: Human Papiloma Virus (HPV) Genital Warts Molecular (PCR) Pap Smear
Z Nazari, E Tabarraei, J Akbarmehr,
Volume 8, Issue 1 (4-2014)
Abstract
Abstract
Background and Objective: Respiratory tract infections (RTI) are the most common infectious disorders, worldwide. About 80%-90% of RTI are caused by four viruses such as Adenoviruses, 51 serotypes have been introduced so far. The aim of this survey was to evaluate the frequency of Adenovirus in respiratory infected patients by PCR method in Golestan province, Iran.
Material and Methods: This descriptive cross-sectional study was conducted on 400 patients with clinical diagnosis of flu-like respiratory infection, 2010-2012. In addition to collecting demographic and clinical data, nasopharyngeal swabs were taken and transferred to the virology laboratory in viral transport medium (VTM), and evaluated by PCR method for Adenovirus after genomic extraction. Using SPSS v.11 software, we analyzed the data.
Results: Thirty-seven (9.2 %) were positive for Adenovirus. No significant correlation was found between being positive for Adenovirus and the variables such as age, gender and season. Clinical signs were coughing (27 73%), body pain (25 67.6%), and fever (24 64.9%). Thirty-five of the patients (94.5%) had at least one symptom.
Conclusion: Our findings are consistent with other research conducted in Iran and other countries. There is a significant correlation between Adenovirus infection and clinical symptoms.
Keywords: Respiratory Infection, Adenovirus, PCR, Golestan, Iran
S Zhand, A Tabaraei, A Moradi, F Fotoohi, N Javid, M Bazoori, E Haji Mohammadi, A Ghaemi,
Volume 8, Issue 2 (7-2014)
Abstract
Abstract
Background and Objective: The emergence of a novel H1N1influenza A virus of animal origin with transmissibility from human to human poses pandemic concern. Current subtypes of Seasonal influenza A viruses spread in human are influenza A H1N1 influenza A H3N2 and influenza type B viruses. The aim of this study was to determine current strains of the H3N2 and new H1N1 subtypes of influenza A virus from patients suspected influenza infection in 2009 flu pandemic in Golestan province, Iran.
Material and Methods: In this descriptive study, respiratory samples (n = 153) from patients with acute respiratory symptoms were collected in 2009 flu pandemic applied during 2009 pandemic influenza in Golestan province. After reverse transcription of extracted viral RNA, PCR was developed for both H1N1and H3N2subtypes using CDC specific primers.
Results: The mean age of patients was 16.59. Of them 45.1% were male. Thirteen (8.49%) were infected with seasonal influenza H1N1 and 25(16.33%) with seasonal H3N2influenza.
Conclusion: The rate of infection with seasonal H1N1and H3N2is similar to other studies reported from Iran, but lower than the rate reported from other parts of the world.
Key Words: Influenza A Virus, H1N1, H3N2, RT-PCR, Iran
F Momeni, R Ghanbarpour, L Dolatshah, H Alizade,
Volume 8, Issue 2 (7-2014)
Abstract
Abstract
Background and Objective: CTX-M type extended spectrum beta-lactamases is a rapidly expanding group of enzymes encountered with increasing frequency, especially, in Escherichia coli (E. coli). There are a few reports on phylogenetic background of E. coli isolates from clinical sources of under five-year- old children in Iran. The purpose of this study was phylotyping of E. coli isolates having blaCTX-M and blaCTX-M-15 genes from under five-year- old children with diarrhea and urinary tract infection (UTI).
Material and Methods: A total of 121 E. coli isolates (75 diarrheas and 46 UTI) were obtained and identified as E. coli based on standard bacteriological tests. DNA was extracted from E. coli isolates by alkaline lysis method. PCR assay was used because of high frequency of blaCTX-M and blaCTX-M-15 genes in the isolates and also determination of phylogenetic group/subgroups by detection of yjaA and chuA genes and fragment TspE4.C2.
Results: The isolates belonged to four phylogenetic groups A (48.77%), B1 (14.04%), B2 (11.57%), and D (25.62%). In the diarrheic isolates,17.37% were positive for blaCTX-M and 14.04% of isolates possessed both blaCTX-M and blaCTX-15genes.Out of 46 UTI isolates, 21.73% were positive for blaCTX-M and 15.21% for blaCTX-M and blaCTX-M-15 genes.
Conclusion: A rather high prevalence of E. coli isolates with blaCTX-M and blaCTX-M-15 genes was observed in fewer than five-year- old children in Khoramabad city. Phylotyping of isolates possessing blaCTX-M and blaCTX-15genes showed that most of them belonge to A and D phylo-groups.
Keywords: Escherichia Coli, Phylogenetic Group, Extended-Spectrum Beta-Lactamase
A Hajihasani, M Douraghi, M Rahbar, M Mohammadzadeh, H Zeraati, S Ghoorchian, M Alavi Moghadam, M Sabzi,
Volume 8, Issue 2 (7-2014)
Abstract
Abstract
Background and Objective: Stenotrphomonas maltophilia is an opportunistic nosocomial pathogen with high mortality in immunocompromised cases. The aim of this study was to isolate and identify Stenotrphomonas maltophilia in the hospitals’ environment and wards.
Material and Methods: In this cross-sectional study, a total of 1108 samples were collected from environment of two hospitals during 12 months. Identification of isolates was performed using biochemical, phenotypic (intrinsic resistance to carbapenems) and molecular methods (amplification of 23S rRNA gene).
Results: Of the studied samples, 186 (16.78%) nonfermentative gram negative bacilli (NFGNB) were identified. Amongst NFGNB, 18 (1.62%) isolates were identified as S. maltophiliaby using biochemical tests. Of 18 biochemically identified isolates, 15 (83.3%) were confirmed via PCR. Sinks (40%) and men surgery ward ( 33.3 %( were the most contaminated sites and wards of hospitals, respectively.
Conclusion: S. maltophilia is repeatedly isolated from sink which shows that the moist hospital environments need to be considered as a source for dissemination of bacteria.
Keywords: Nosocomial Infections, Nonfermentative Gram Negative, Stenotrphomonas Maltophilia, PCR
M Hasannejad Bibalan, N Javid, M Samet, F Shakeri, Ea Ghaemi,
Volume 8, Issue 3 (8-2014)
Abstract
Abstract
Background and Objective: Biofilm is a complex microbial community embedded in a self-produced extracellular polymeric matrix. We aimed to study the extent of biofilm formation by S. Areas isolates and its relation to some phenotypic and genotypic criteria.
Material and Methods: One hundred-fifty strains of Staphylococcus aureus isolated from Gorgan were studied. Microtiter plate assay method was used for investigation of biofilm formation.The biofilm formation of strains were recorded and its relation to accessory gene regulator (agr) and antibiotic resistance were assessed by X2 test.
Results: Eighty-four isolates (56%) were able to form biofilm. The strength of biofilm formation in agr group I was more than that of other groups. The biofilm formation among S. Areas isolated from the wound and urine (both with 75 %) had the highest capability. Methicillin-resistant isolates had a greater ability to biofilm formation.
Conclusion: Methicillin resistant isolates had a greater ability to biofilm formation. Given the importance and treatment related problems of Methicillin-Resistant Staphylococcus Aureus (MRSA) especially Community Acquired-Methicillin-Resistant Staphylococcus Aureus (CA-MRSA), it is a necessity to control or remove the biofilm formation alongside antibiotic treatment.
Keywords: Staphylococcus Aureus, Biofilm, Microtiter Plates Assay, PCR
H Ghaffari, A Moradi, A Ghaemi, N Javid, M Talkhabifard, H Naziri, A Tabaraei,
Volume 8, Issue 3 (8-2014)
Abstract
Background and Objective: Cytomegalovirus (CMV), one of the most common opportunistic pathogens in patients infected with human immunodeficiency virus (HIV), can cause the diseases such as encephalitis, pneumonia, and chorioretinitis. This study aimed at molecular studying of CMV infection in individuals infected with the human immunodeficiency virus. Material and Methods: In this study, 50 venous blood samples from HIV-infected individuals were taken. Patients were divided into two categories: patients under treatment with and without antiretroviral drugs. Plasma were separated from blood samples and examined for the presence of cytomegalovirus genome by PCR. Material and Methods: this study was conducted on 50 blood samples from HIV-infected individuals, and plasma was separated and examined for the presence of cytomegalovirus genome by PCR. Patients were divided into two group of under treatment with and without antiretroviral drugs. Results: Of 50, 28 (% 56) were men and 22 (% 44) were women. CMV genome was identified in 8 samples (16%), and the molecular prevalence of CMV infection was 21.4% (n= 6) in males and 9.1% (n = 2) in females. Conclusion: Given the frequency of Cytomegalovirus Active Infection in HIV-infected individuals under antiretroviral therapy, we should be careful about the treatment of Cytomegalovirus Active Infection. Keywords: Active Infection, Cytomegalovirus, Human Immunodeficiency Virus, Shiraz, PCR
Zare, Z, Solhjoo, K, Norooznejad, Mj, Kazemi, A,
Volume 8, Issue 4 (1-2015)
Abstract
Abstract
Background and Objective: Increasing prevalence of methicillin resistant Staphylococcus aureus strains (MRSA) with their multidrug resistance potential causes difficulties in the treatment of infections due to these bacteria. Hence, the detection and determination of the frequency of MRSA strains via phenotypical and molecular methods is necessary in different parts of the county.
Material and Methods: In this cross- sectional study, 150 Staphylococcus aureus strains were collected from different clinical samples in the hospitals located in Shiraz and Jahrom, Iran. To detect methicillin resistant Staphylococcus aureus strains, we used phenotypical methods such as disc diffusion and minimum inhibitory concentration by E-Test, and PCR molecular method for mass gene.
Results: The frequency of methicillin resistant Staphylococcus aureus was 63 strains (42%) using disc diffusion and E-Test. while in PCR method, in addition to 63 strains, nine other isolates, which were sensitive to oxacillin by disc diffusion and E-Test, possessed also mecA gene. By and large, 72 isolates (48%) had methicillin resistance gene.
Conclusion: Given the results of phenotypical and molecular methods, the frequency of methicillin resistant Staphylococcus aureus was relatively high in this area. Thus, the MRSA strains can be detectable as soon as possible by accurate and sensitive methods such as PCR to determinate the effective antibiotics.
Keywords: Methicillin Resistant Staphylococcus Aureus, MRSA, MecA Gene, PCR
M Talkhabifard, M, N Javid, N, A Moradi, A, A Ghaemi, A, A Tabarraei, A,
Volume 8, Issue 5 (1-2015)
Abstract
Abstract Background and Objective: Human Cytomegalovirus (CMV) is an important cause of congenital viral infection that can lead to serious diseases and complications in infants. Application of rapid, sensitive, and specific HCMV detection methods is necessary for congenital infection detection. We aimed to optimize the use of PCR and ELISA for detection of HCMV in infants. Material and Methods: PCR–ELISA was performed by using specific primers and probe for detection of the HCMV glycoprotein B gene. First, the extracted DNA from urine samples and controls were labeled by digoxigenin during DIG-labeling PCR. After that, Biotin-labeled probe captured the DIG-labeled PCR products. The probe-PCR product hybrid is immobilized on a streptavidin-coated Microtiter plate, and detection was confirmed by proxidase-conjugated anti-digoxigenin antibody, and calorimetric substrate. Results: The clinical Human CMV strains isolated from16 patients were detected by this method. The optimized PCR-ELISA method was able to detect less than100 copies of HCMV genome. There was no non-specific reaction. Conclusion: PCR-ELISA can be applied as a sensitive, specific and reliable method for Semi-quantitative CMV detection in clinical samples. Keywords: Cytomegalovirus, Glycoprotein B, PCR-ELISA, Semi-Quantitative
Nasrollahi Omran, A, Nazemi, A, Kihanian, Sh, Aryana , N,
Volume 8, Issue 5 (1-2015)
Abstract
Abstract Background and Objective: With the development of drug resistance in strains of fungi, there is a considerable resistance of Candida albicans strains to fluconazole. Molecular studies are developing to determine the relationship of such a drug resistance with the increased gene expression of enzymes produced in drug-resistant Candida isolates. We aimed to evaluate the relationship between extracellular lipase gene (LIP8) expression of Candida albicans isolated from candidiasis and sensitivity or resistance to fluconazole. Material and Methods: Drug susceptibility of Candida albicans was performed in oral and vaginal candidiasis to determine the proportion of strains sensitive or resistant to fluconazole using NCCLS method. To evaluate and compare the expression of these genes in the susceptible and resistant strains, RT real-time PCR reaction was used. Results: Of 46 Candida albicans, 20 were susceptible, 12 were semi-susceptible and 14 were resistant to fluconazole. By using PCR reaction, the results showed that the expression of this gene in fluconazole-susceptible isolates was moderate, while it was high in the isolates resistant to fluconazole. Conclusion: The results of lipase gene (LIP8) expression showed that the additional expression of some genes of the enzymes responsible for virulence of Candida may also play a role in resistance to fluconazole. Keywords: Candidiasis, Lipase Gene Expression, RT real-time PCR, Fluconazole
