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Showing 109 results for Pcr

H Bagheri, A Ghaemi, M Aslani, N Mozafari, S Livani, T Dadgar,
Volume 2, Issue 1 (4-2008)
Abstract

Abstract Background and objectives: Diarrhea is one of the main cases of morbidity and mortality among children in developing countries. Enteroaggregative Escherichia coli (EAggEC) is an emerging diarrheal pathogen that has been associated characteristically with persistent diarrhea among infants, particularly in the developing Counties. Therefore, we decided to study the prevalence of enteroaggregative strain in cases of Diarrhea in Gorgan by PCR method. Material and Methods: This descriptive study was carried out on 455 subjects suffered from Diarrhea in Gorgan during one year (2005-6). At first, the samples were cultivated on the MacConkey agar and EMB agar media, Then all colony Suspected to E.coli were chosen and their DNA extracted by phenol chloroform method. The result was obtained by the selected primer, PCR method. Results: of 455 samples, Twenty cases (4/4%) including men (12) and woman (8) are positive for EAggEC, 85% of sufferers are under 5 years old (45.8% of them are under one year old). The Prevalence of this gene in Summer , Autumn ,Winter ,Spring are 5.3% , 4.2% , 4.1% and 1.8% ,respectively. Conclusion: Based on the prevalence of Enteroaggregative Escherichia coli (EAggEC) in Diarrheagenic cases in Gorgan (4.4%), we do recommend using molecular methods, which are reliable and less expensive than classic methods, in detecting of microorganisms. Key words: Entroaggregative Escherichia coli, Diarrhea, PCR, Gorgan.
Hr Tavakoli, M Manafi, M Bayat, A Mehrabi Tavana,
Volume 2, Issue 2 (10-2008)
Abstract

Abstract Background and Objectives: Chromogenic media are the newest methods applied for rapid detection of pathogenic microorganisms in drinking water and food from 1998-2008. These Specific media contained the compounds acting as a substrate for microbial enzymes and, according to the type of enzyme, produce specific color. The aim of this study was to introduce the chromogenic media as a powerful tool in rapid detection of pathogenic agents in drinking water and food. Material and Methods: In this review article, the published papers about the use of chromogenic media in rapid detection of water and food-born pathogens were investigated. Results: The studies conducted in different countries show that the chromogenic media are very sensitive, specific and with high performance therefore, we can use it to detect the most important pathogenic microorganisms (such as Salmonella spp, E.coli, S.aureus, L.monocytogenes, and Candida spp.) in water and food samples. Conclusion: Because chromogenic media, in comparison with the other rapid detection methods such as PCR and ELISA, are very sensitive and cheaper, it can be applied as an alternative method. Key words: Chromogenic media, Rapid detection, water and food, Microorganism
N Ziaei, N Amir Mozafari, H Kouhsari, A Moradi, A Tabarai, T Dadgar, S Livani, M Arab Ahmadi,
Volume 2, Issue 2 (10-2008)
Abstract

Abstract Background and Objectives: Diarrhea is one of the most common diseases in the world. Campylobacter jejuni is one of the prevalent agents of bacterial diarrhea in most of developing countries. It is usually ignored in routine laboratory test in our country, because it has a difficult investigation method. This article aims to determine the prevalence of Campylobacter jejuni, in diarrhea samples in Gorgan City (East north of Iran) by PCR Method. Material and Methods: This descriptive study was performed on 455 diarrheal samples during one year (2005-06).255 out of them were cultured on Preston media (Himedia co.) on 42°c. DNA Extracted by phenol cholorophorm method was directly carried out on stool samples.16srDNA hipo and asp primers for detection of Campylobacter genus, C.jejuni and C.coli species were used, respectively. In addition, universal primer of 16srDNA was used for control of PCR method. Results: no sample was positive for Campylobacter in culture .only three samples were positive for Campylobacter genus and C.jejuni specific primer but none of them were positive for C.coli .99 samples were positive by universal primer of 16srDNA . Conclusion: The results indicate that C.jejuni isn't a prevalent agent in diarrhea in our region. Key words: Campylobacter jejuni -Gorgan- Diarrhea
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
F Shrafati-Chaleshtori, R Sharafati-Chaleshtori, A Shakerian, H Momtaz,
Volume 3, Issue 1 (4-2009)
Abstract

Abstract Background and objectives: Paratuberculosis or John's disease is a chronic infectious disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). It results in major economic losses to dairy farm of all over the world and it is the agent causing crohn's disease. The aim of this study was to detect the MAP using PCR in raw-milk samples of cows in shahre-kord. Material and Methods: In this cross–sectional study, 100 raw milk samples of cows were collected from both industrial and semi -industrial farms in shahre-kord. The DNA of all Samples was isolated by MAP, using PCR method. Results: The results Show that only three (3%) Samples were positive for Mycobacterium avium subsp. paratuberculosis. Conclusion: Based on our results, Milk -PCR was useful for detection of MAP in milk samples. Key words: Mycobacterium paratuberculosis, milk, polymerase chain reaction.
A Moradi, E Mobasheri, A Tabarraei, S Bakhshandeh Nosrat, V Kazemi Nejad, R Azarhosh, Sh Alizadeh, M Bazori,
Volume 3, Issue 1 (4-2009)
Abstract

Abstract Background and objectives: Breast cancer is the most prevalent one in women. Some of the common causative factors are genetic background, nutritional and environmental factors. Viruses are believed as a risk factor in this cancer, too. Recent studies reported that Human Papillomaviruses can be one of the possible risk factors of breast cancer. This study focused on investigation of the papillomavirus genome in tissues of breast cancer in Golestan province, Iran. Material and Methods: This descriptive analytical study was done from 2005 until 2008. The Samples were obtained from women admitted to the hospitals in Gorgan and Gonbad cities. All breast biopsy or mastectomy tissues were confirmed by the pathologists for breast cancer. DNA was extracted and PCR done by using general primers (GP5 + / GP6 + and MY09/MY11) for detection of papillomavirus genomes. Results: The Subjects are 231 patients aged 47± 12/72, the youngest 20 and the oldest 84. They are from Gorgan (N=122)and Gonbad (N=109) The result Shows That The Subjects Suffer from infiltrating ductular Carcinoma(31.4%), infiltrating duct Carcinoma (60.1%)and intraductal Lobular Carcinoma (4.3%) and The rest from other kinds of Cancer. Papilloma Virus genome is not found in These Samples. Conclusion: Based on paradoxical results from different parts of the world, upon the presence or absence of papillomavirus genome in breast cancer samples, to show the role of this virus in the development of breast cancer more studies are needed. Key words: Breast Cancer, Papillomaviruses, Golestan Province
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
F Ghasemi Kebria, B Khodabakhshi, H Kouhsari, M Sadeghi Sheshpoli, N Behnampoor, S Livani, M Bazuori, E A Ghaemi,
Volume 4, Issue 1 (4-2010)
Abstract

Abstract Background and objectives: After respiratory infection, Diarrhea is the second cause of mortality. Yersinia enterocolitica is the second important cause of infectious diarrhea in children of some countries. In this study, we evaluated the frequency of Yersinia entocolitica of diarrheal specimens in Gorgan, Iran. Material and Methods: This descriptive cross - sectional Study was carried out on diarrheal stools of 455 patients referred to medical centers and laboratory of Gorgan in 2004-2005. DNA extraction using phenol chloroform was performed for all samples. Using two specific primers (genus-specific16s rRNA and ail- specific species genus of Yersinia enterocolitica), we did PCR sample. Results: Yersinia genome was identified in 12 patients(2.63%) and 11 of them was Yersinia enterocolitica. The frequency infection in of girls (3%) was more than boys (2.4%), and the prevalence in winter (4%) was more them other seasons, and under one- year- group (3.4%) and 1-5 years (3.1%) is more than other age groups. It was not observed significant difference. (P> 0.05). Conclusion: The frequency of Yersinia in cases of diarrhea in Gorgan is similar to most regions of Iran and in children under 5 years is observed more in winter. Key words: Yersinia enterocolitica, Diarrhea, children, Gorgan
Mahsa Yazdi, Ali Nazemi, Mir Saed Mir Nargasi, Mr Khataminejad, Sh Sharifi, M Babai Kochkaksaraei,
Volume 4, Issue 1 (4-2010)
Abstract

Abstract Background and objectives: Beta-lactamase enzymes are the most causes of resistance to antibiotics among gram-negative bacteria. Nowadays, Infections due to ESBLs are being increased throughout the world and is considered as a new burden to the health systems. This study aimed at determining the sensitivity pattern of E.coli isolates to beta-lactam antibiotics, and investigating the presence of blaCTX-M, blaTEM, and blaSHV genes in the urine samples. . Material and Methods: In this study, 244 E.coli isolates were screened in 2009-2010. The antibiotic susceptibility of E. coli isolates were determined by disc-diffusion method. Antimicrobial agents tested were cefoxatime, ceftazidime, imipenem, nalidixic acid, and ciprofloxacin. The combined disc test was used to confirm the results. The results were compared to the Clinical and Laboratory Standards Institute (CLSI) and ESBL positive isolates were further investigated for the presence of blaCTX-M, blaTEM, and blaSHV genes by PCR. Results: Of 244 E. coli isolates, 116 (47.1%) are resistant to Ceftazidime, and 96 (39.2%) to cefoxatime. Also, 109 (44.3%) isolates are ESBL positive. blaCTX-M, blaTEM, and blaSHV genes are found among 95 (87.1%), 75 (68.8%), and 77 (70.6%) ESBL positive isolates, respectively. Forty (36.6%) isolates have all three genes, while 68 (62.3%) include blaTEM and blaSHV genes. Moreover, 61 (55.9%) isolates carry blaCTX-M and blaTEM genes, and 54(49.5%) have blactx-M and blashv. Conclusion: Regarding the high frequency of resistance to the third generation cephalosporin antibiotics, precise antibiogram testing is highly recommended before any antibiotic prescription in cases of infections with ESBL producing microorganisms. Key words: ESBL Escherichia coli blaCTX-M blaTEM blaSHV.
A Nazemi, M Naderi, M Jafarpour, M Mirinargesi, Sh Sharifi,
Volume 4, Issue 2 (10-2010)
Abstract

Abstract Bachground and objectives: The ability of adherence to the surface of host cell is very critical in the colonization of microbial pathogens. It has been revealed that E. coli strains that infect urinary tracts have different fimbrea such as I, S, FIC, Dr, and fimbrial adhesions. Material and Methods: In this study, 363 urine samples were obtained from patients with urinary tract infections reffered to clinical laboratories in Western areas of Tehran ,2008-2010 by using biochemical tests,200 samples were confirmed to be E.coli.First, DNA was extracted by boiling method and then the presence of fimbria fim, sfa, pap, foc, and afa genes tested by PCR. Results: In 200 samples, the frequency of fimbria fim, sfa, pap, foc, and afa genes are188 (%94 ), 34 (%17), 20 (%10), 61 (%31) and 71 (%35.5), respectively. Conclusion: The resultes show that FIM ans SFA are the most fimbrial genes of E. coli isolated from urine samples .This information can be valuable in etiology of urinary tract infection (UTI), UTI administration, and making of new vaccines. Key words: Urinary tract infection, fimbria, Uropathogenic E. coli (UPEC)
S M Hashemi, A Nasrollahi Omra, Hr Pordeli, A Hosenian,
Volume 4, Issue 2 (10-2010)
Abstract

Abstract Bachground and objectives:Streptomyces is the most important genus in Actinomycetes family.The Streptomycetes are widely used in industry producing numerous chemical compounds including antibiotics, enzymes and anti- tumor agents. The aim of this study was to isolate soil-borne Streptomyces producing antimicrobial substances from soil of Golestan province of Iran and to survey anti-fungal metabolities produced by this organism. Material and Methods:In this study various soil samples were collected: ( forest areas of Naharkhoran in Gorgan and Kordkuy’s Derazno, Aghala’s deserts and agriculture lands of Aliabad ) were cultured on Actinomycet isolation agar and Starch casein agar were identified and purified by morphology and biochemistry tests. The activity of isolated Streptomyses against:Aspergillus flavus, Aspergillus, Candida albicans and Malasesia fur fur were studied by Agar Diffusion. Results:Of 120 samples, 24 are Streptomyces(20%) .The frequency of Streptomyces are reported in Aghala (10,41.6%),Derzno (8,33.3%) ,Nahar khoran(4,16.6%) and Aliabad(2,8.3%).Of 24 isolated Sterptomyses,two isolates have strong anti-fungal and six of them have moderate effect.We also see Streptomyses,isolated from desert area, have higher anti-fungal activity. Conclusion:It is recommended two isolated of Streptomyses be identified ana purified. Key words:Streptomyces , antifungal activities, antibiotic


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
Jafarpur M, Nazemi A, Mirzaee A, Rahbar Farzamee Hagh S,
Volume 5, Issue 2 (10-2011)
Abstract

Abstract Background and objectives: Group A Streptococcus (GAS) strains have been identified by serologic methods based on surface protein antigens, T and M. Accordingly, different serotypes have been reported worldwide. Recently, the previous out of date procedures have been replaced by N-terminal emm gene sequence, which has been used in identifying more than 150 emm types. We aimed to determine the prevalence of emm types and phenotypes resistance to erythromycin among streptococci isolated from the throat in north of Iran. Material and Methods: 50 GAS isolates from sore throat of patients referred to a few local hospitals in Tonekabon, Ramsar, and Chalus in northwest of Iran (2010-2011), by using blood agar, bacitracin sensitivity test, PYR test and agglutination by specific antiserum. Antibiotic resistance of the isolates was determined by the discs branded by Iranian Padtan Teb Company, using Kirby Bauer Test, and analyzed by CLSI standards. The mechanism of resistance to erythromycin was evaluated by Double Disk Diffusion Test in the presence of erythromycin and clindamycin. emm gene of all isolates were reproduced and their PCR products sequenced by the Korean Macrogen company. To determine the emm types, using BLAST2.0 program (National Center for Biotechnology Information, available in WWW.ncbi.nlm.nih.gov / BLAST), and the emm gene sequences were compared with sequences in the gene bank. Results: we identified Four different types of emm, including e mm5 (26 52 %), emm12 (12 24%), emm79 (6 % 12) and emm86 (6 % 12). All beta lactam antibiotics have inhibitory effect on isolates, while18% of isolates (9 of 50) are resistant to erythromycin. The most common resistance phenotype is cMLSB (% 66.6) and the next one is phenotype M (% 33.3), but phenotype iMLSB is not observed in none of the isolates. Twelve percent (6cases) of isolates are resistant to clindamycin. Conclusion: The results of present study show different types of GAS than those reported worldwide. The emergences of emm86 in pharyngitis and erythromycin resistance are the two valuable findings of this research. Keywords:Streotococcus pyogenes,erythromycin,cMLSB,iMLSB
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
Soltan Dallal. M.m, Rahimi Forushani, A., Sadigh Maroufi, S, Sharifi Yazdi, K,
Volume 5, Issue 2 (10-2011)
Abstract

Abstract Bachground and objectives: Salmonella is one of the most important agents of gastrointestinal infection and diarrhea in our country. Misdiagnosis of these bacteria leads to cure failure. The aim of this study was to make a comparison between PCR and the API-20E and conventional biochemical tests carried out for the identification of Salmonella. Material and Methods: In this study 470 specimens taken from children, with acute gastroenteritis, referred to teaching hospitals called Imam, Shariati and children medical centre. The specimens were transferred to microbiology laboratory in public health school for identification of Salmonella with PCR and API-20E methods. Results: Of 470 specimens, 65(13.8%) are positive for salmonella in hospital laboratory, while 37 (7.9%) for API-20E and 39 (8.3%) for PCR are positive. The results of antibiotic sensitivity tests on 39 salmonella isolated from diarrhea specimens show that 73.3% of them are resistance to at least one of the sixteen antibiotics tested. Conclusion: Based on the the results, there is significant difference (P<0.05) between conventional method, API-20E and PCR Key words: Salmonella, conventional identification, molecular identification
Tajeddin, E. (msc), Jahani Sherafat, S. (msc), Seyyed Majidi, M. R. (md), Alebouyeh, M. (phd), Nazemalhosseini Mojarad, E. (msc), Pourhossengholi, A. (phd), Mohammad Alizadeh A H (md), Zali, Mr (md),
Volume 5, Issue 2 (10-2011)
Abstract

Abstract Background and objectives: Bile in healthy people is a sterile fluid and presence of any microorganism can be a marker for a disorder like cholelithiasis. The aim of this study was to determine the frequency of bacterial agents in the bile of patients with bilestone, malignant pancreatic and biliary diseases. Material and Methods: One hundred and two bile samples were obtained, during six months in 2011, from patients subjected to ERCP in Taleghani hospital, Tehran. First, Patient's clinical data, the type stone, and their disease status were studied, and then the microbiological investigations, such as culture, identification of the bacteria and detection of their counts, drug susceptibility testing and molecular tests (16s rDNA PCR) performed on all the samples. Higher than 103 bacteria counts for each sample, in the absence of underlying infections, was considered as stable colonization. We run SPSS version 13 to analyze the data. Results: Out of 42(41.1%) positive bile culture samples, 59 bacterial isolates are detected by conventional methods. Of culture negative samples, seven have bacterial DNA indicated by PCR method. The most isolated bacteria are E. coli (%34.4), Enterococcus spp. (%19.7), Klebsiella pneumoniae (%18) and Pseudomonas aeruginos (18%). The most frequent stones are cholesterol, black pigment and brown pigment, respectively. There is no significant association between the diseases, stones and types of bacteria. Previous antibiotic usage (44.6%) is meaningfully more than that of other biliary problems (p=0.01) Conclusion: The presence of bacteria, Escherchi coli and Entrococcus which are the most in bile samples, is considered as a risk factor in pathogenesis of biliary disorders. Further studies on the pathogenesis and pathophysiological effects of bacteria can help us to clarify the role of bacteria in producing bile stones. Key words: Bile stones, Bacteria, ERCP, Antibiotics.
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

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