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Articles by B. Kazemi
Total Records ( 13 ) for B. Kazemi
  M.R. Roozbahani , M. Bandehpour , A. Haghighi- Khiabanian-Asl , H. Abdollahi and B. Kazemi
  The aim of this study was designing a diagnostic kit for yersiniosis in the trout fish in Iran. Colonies of Yersinia ruckeri were collected from culture medium and a suspension was prepared in a lysing solution. DNA was extracted through a boiling and phenol chloroform method. Two primer sets targeting bacterial 16S rRNA and trout 18S rRNA. Polymerase chain reaction products were separated by gel electrophoresis. Among 20 suspected samples tested two samples were positive for both host and bacterial PCRs indicating the positive Y. ruckeri infection and remaining 18 samples were negative for pathogen. The performance of PCR reactions in negative samples were confirmed from amplification of internal control reactions targeting host. A PCR based diagnostic kit with an internal control was prepared for detection of Yersinia ruckeri in rainbow trout fish.
  M. Afsharnasab , R. Mortezaei , V. Yegane and B. Kazemi
  The importation of Litopenaeus vannamei to Iran from Hawaii was initiated when Iranian shrimp culture was first affected by WSSV in 2004. The main reason for the importation of L. vannamei to Iran was the disease susceptibility and mass mortality of the indigenous species (P. indicus) when faced with the first outbreak of WSSV. During the two years of study, it was found out that culturists in Iran preferred cultured L. vannamei than the local species (P. indicus). In 2008, mass mortality occurred in farmed L. vannamei in Khuzestan Province South of Iran. Two hundred shrimps with white spot on the carapace and body were collected and preserved in Davidson fixative for histopathology. A part of samples collected were also preserved in 95% ethyl alcohol for Polymerase Chain Reaction (PCR) technique. Two pair primers from VP24 WSSV genome was identified and used for PCR while identified one pair primer for 18SrRNA gene of shrimp was used as house keeping gene in PCR reaction in both positive and negative PCR reaction. Grossly, the samples showed white spot in the cuticle and body surface and red color on the appendages. Histopathologically, all tissue except hepatopancreas showed the intranuclear Cowdry type-A inclusion bodies. PCR studies using designated primer revealed a band of 414 bp from WSSV and 809 bp of shrimp DNA fragments in positive samples. The negative samples showed just 809 bp. This is the first report of White Spot Syndrome Virus (WSSV) in farmed L. vannamei in Iran.
  P. Ghadam , F. Sadeghian , R. Sharifian , S. Sadrai , B. Kazemi and E. Nematipour
  The vitamin K epoxide reductase subunit 1 (VKORC1) has been identified recently. It is a component of the enzyme vitamin K epoxide reductase that is the therapeutic target site of warfarin. In order to investigate the relationship between VKORC1 gene and warfarin dose response, we studied this gene in an Iranian warfarin resistant patient who receive more than 100 mg warfarin per day. The results showed that although warfarin concentration in his plasma was extremely higher than therapeutic level (22.8 mg L-1) but no mutation(s) found in the exons of VKORC1 gene. Other genes may be contributed in resistance to warfarin in this patient.
  N. Amirmozafari , R. Mirnejad , B. Kazemi , E. Sariri , M.R. Bojari and F. Dehdar Darkahi
  The aim of this investigation was to simultaneously detect Mycoplasma hominis, Mycoplasma genitalium and Ureaplasma urealyticum in female patients suffering from genital complications (vaginitis, cervicitis and PID) by PCR. Genital swabs were collected from 210 female patients and subsequently suspended in PBS. Following DNA extraction, PCR assay were performed, using a genus specific primer pair. These primer set, which were originally designed in our laboratory, amplified a 465 bp fragment (M. genitalium), 559 bp fragments (U. urealyticum) and 630 bp fragment (M. hominis). Samples containing bands of the expected size for mycoplasma strains were subjected to digestion with restriction endonuclease enzyme. Of the 210 genital swabs tested, 120 sample (57.1%) showed positive reactions in PCR. Sixty eight samples were positive for Ureaplasma sp. (32.3%), 28 for Mycoplasma sp. (13.3%) and 25 samples had mixed infections (11.9%). In case where specific primers were utilized, PCR has proved to be a simple, fast and relatively inexpensive method for simultaneous detection of all three clinically important genital mycoplasmas.
  M. Norouzi , M. Pourkazemi , A. Keyvan , S.M.R. Fatemi and B. Kazemi
  In this study, 197 samples of adult stellate sturgeon from four fishery regions were collected. DNA was extracted using 15 pairs of microsatellite primers, Polymerase Chain Reaction (PCR) was conducted. DNA bands were analyzed using Biocapt and GenAlex software package. Out of 15 microsatellite primers, 11 loci were produced, in which 10 of them were polymorphic and 1 monomorph. Analyses revealed that average of 13 alleles per locus (range 8 to 18 alleles per locus). Average observed and expected heterozygosity were 0.650 and 0.855 and significant genetic differences between 4 regions were observed (p≤0.01). Deviations from Hardy-Weinberg equilibrium were in most cases. Maximum genetic difference were observed between regions 2 and 4 (FST = 0.063, Nm = 3.728). These results indicate that at least, 3 populations of stellate sturgeon exist in the South Caspian Sea. Population of stellate sturgeon in region 2 where Sefidrud drainage is located was consider as independent population, therefore management of this unique stocks for restocking and conservation of gene pools is highly recommended.
  B. Kazemi , Bandehpour , H. Yahya zadeh , M. Roozbehi , N. Seyed , A. Ghotasloo and A. Taherpour
  HCV is one of the main causes of chronic hepatitis in developing countries and is particularly distributed via blood and blood product transfusion. ELISA serological method is currently used as laboratory diagnostic test in Iran, but both clinical and laboratory diagnosis of acute HCV infection remains yet as a problem. The aim of this study was to determine results obtained by PCR and ELISA techniques in detection of HCV in serum samples. Using ELISA and RT-PCR, 159 serum samples were analyzed. Thirty four negative samples (22.35%) and 45 positive samples (27.95%) were detected by both techniques. Thirty two samples (19.88%) were positive by PCR and negative by ELISA and 48 samples (29.82%) were negative by PCR and positive by ELISA. Present results show that there are significant differences between PCR and ELISA diagnostic tests for HCV detection.
  B. Kazemi , F. Yasaee , M. Bandehpour , N. Seyed , Y. Mehrabi , M. Rajabnejad , M. Mansouri , S. Givrad , M. Ghazi and H Baseri
  This study was conducted for parasite detection in urine samples using Polymerase Chain Reaction (PCR) and comparing the results to wet mount microscopic screening. For this, 155 urine samples were collected from symptomatic women suspected to trichomoniasis (mean age = 20±5 year) introduced to laboratory by physician. Urine samples were subjected to polymerase chain reaction to detect the parasite's genome. As a result 25 out of 155 samples (16%) were detected positive by direct microscopic observation, but 75 out of 155 (48%) by PCR. Eighty out of 155 samples (52%) were detected negative by PCR. This study confirmed that the microscopic screening with a low sensitivity must be substituted by highly sensitive screening methods such as PCR.
  A. Haghighi , S. Rasti , E. Nazemalhosseini Mojarad , B. Kazemi , M. Bandehpour , Z. Nochi , H. Hooshyar and M. Rezaian
  The nucleotide sequences of Serine-Rich Entamoeba histolytica Protein (SREHP) gene have already exhibited stable and significant polymorphism in the gene studies. Serine-rich protein is also present and polymorphic in Entamoeba dispar which called SREDP. The polymorphism of the Serine-Rich Entamoeba dispar Protein (SREDP) gene among 8 isolates obtained from Iranian cyst carriers were analyzed by a nested PCR-RFLP followed by sequencing of the PCR products. From those isolates, six distinct DNA patterns were observed after PCR-RFLP of the nested PCR, whereas sequencing showed 8 different patterns among the isolates. The results demonstrate an extensive genetic variability among Iranian E. dispar isolates. The repeat-containing region of the SREDP was found extensively polymorphic in size, number and order of repeat units. Genetic diversity of Iranian E. dispar isolates based on the SREDP was more polymorphic in comparison of Serine-Rich Entamoeba histolytica Protein (SREHP) of the E. histolytica isolates as well as were different from a few known SREDP genes.
  A.M. Saberi , M. Bandehpour , M. Afsharnasab , E. Ghayour , S.A. Yousefi Namin and B. Kazemi
  The aim of this study is designing a diagnostic kit for white spot syndrome virus. We designed 2 series of primers for diagnosis of viral VP24 gene and also primers as internal controls which amplify part of genome in both positive and negative samples. DNA of shrimps were extracted and PCR amplification carried out. In this research, a diagnosis kit for white spot disease of shrimps designed and tested using 32 shrimp samples which were dubious to have this disease. White spot virus were found in 23 samples and the other 9 were negative. For extra confirmation, the PCR product was sequenced and deposited to GenBank. We designed a diagnosis kit for white spot disease of shrimps and tested successfully.
  A. Haghighi-Khiabanian Asl , M. Azizzadeh , M. Bandehpour , Z. Sharifnia and B. Kazemi
  In this research, we confirmed the positive SVC in three Indian carps spp. (1) Rohu (Labeo rohita), (2) Merigal (Cirrhinus merigala) and (3) Catla (Catla catla) with typical histopathologic signs and PCR sequencing. Nested-PCR used to amplify a fragment of viral glycoprotein gene in Shahid Beheshti University M.C. and PCR product was purified and submitted to sequencing and deposited to GenBank at accession No. FJ711168. Two sites in this research that, were placed in Khuzestan Province (Ahvaz City, South of Iran) and Gilan Province (Astaneh Ashrafieh City, North of Iran) aquaculture farms. Samples were prepared for PCR method in both sites (30 pieces from Khuzestan and 30 pieces from Gilan Province), that all of the (100%) samples were positive in Nested-PCR. In addition, other 60 samples for histopathologic studies in both sites (30 pieces from Khuzestan and 30 pieces from Gilan Province), (according to 10% prevalence). All of the (100%) typically samples were with major histopathologic signs. The results of this study indicate that SVC infection can be found in some Indian carp in Iran.
  P. Ghadam , S. Gharavi , F. Yarian , M.R. Soudi , B. Kazemi and M. Bandehpour
  Among some Bacillus species, a protein highly homologous to HU, classified HB and coded by hbs gene. According to the recent studies, the sequence of hbs gene just in one strain of Bacillus subtilis exists in gene bank (ATCC 23857). In this study, DNA from Bacillus subtilis ATCC 6633 was extracted and investigated by PCR. The PCR product was sequenced and shown to differ in just one nucleotide from B. subtilis ATCC 23857. Hence, it was chosen as reference and for the first time, used for non-radioactive labeled probe preparation. The PCR product in Bacillus subtilis with ATCC 6633 was labeled using non-radioactive DIG-labeled nucleotides and conditions of probe preparation and hybridization were optimized and checked it by Southern blotting.
  Z. Kianmehr , P. Ghadam , S. Sadrai , B. Kazemi and R.A. Sharifian
  VKORC1 is a component of the enzyme that is the therapeutic target site of warfarin. In order to investigate the relationship between VKORC1 gene and warfarin dose response, we studied this gene in 22 clinically sensitive patients and 36 clinically normal patients as control group that in previous studies their blood warfarin levels were determined by HPLC and genotyped for CYP2C9. In the majority of these patients, the results had shown that the clinical phenotype was according to the CYP2C9 genotype, but there were sensitive patients with normal CYP2C9 genotype. So in this study for investigation of another reason of sensitivity to warfarin in these patients, VKORC1 was chosen. In order to determine the impact of VKORC1 1173C>T and 3730G>A polymorphisms in warfarin dose variability, a protocol of RFLP based PCR with HinfI and SsiI was used. Polymorphisms of VKORC1 were effective factors in sensitivity to warfarin in the majority of them. However, in some of sensitive patients, there was inconformity of clinical phenotypes with CYP2C9 and VKORC1 genotypes. In conclusion, our findings suggest that CYP2C9 and VKORC1 are not the only effective factors in sensitivity to warfarin and more studies are necessary for investigation of sensitivity reasons in these patients.
  B. Kazemi , F. Fallahian , N. Seyed , M. Bandepour , A. Shabani and P. Ghadam
  Streptokinase, a bacterial protein produced by various strains of haemolytic streptococci, is widely used to treat humans with thrombolytic disease. This protein is antigenic and anti- streptokinase antibodies (Abs) cause allergic reactions and neutralize streptokinase therapeutic effects. To produce an engineered variant of streptokinase being functional and less antigenic than the native molecule. In this study the C-terminal immunodoinant epitope of native molecule was eliminated by Polymerase Chain Reaction (PCR) using specific internal primers. Then PCR-amplified sequence was cloned into pGEMEX-1 expression vector.
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