Showing 49 results for Pcr
S. Amini Kafi-Abad , A. Talebian , F. Ranjbar Kermani , M. Moghtadaie , M. Sobhani , Sh. Samie ,
Volume 2, Issue 3 (3-2005)
Abstract
Abstract
Background and Objectives
Screening the blood donors for serological markers reduced the incidence of transfusion-transmitted infections especially post-transfusion hepatitis C. However, there remains residual risk due to pre-seroconversion period. HCV RNA (PCR) of blood donations reduced the residual risk of transfusion-transmitted HCV infection. In this study, blood donations were screened for HCV RNA by RT-PCR method.
Materials and Methods
An extra plasma sample was collected from 1026 blood donors. 1000 out of 1026 samples were negative for HBsAg, anti-HCV (EIA, third generation), anti-HIV and RPR. Every 5 samples were pooled. The sensitivity of HCV-RNA detection by RT-PCR method was 380 geq/ml according to Proficiency VQC panel. 1000 donations in 200 pools were tested.
Results
False reactivity of samples considered positive accounts for 5.5% of cases, and 5.5% were invalid due to non-specilic bands. 6% of the pools were false-positive. A false positive result was defined as positive on initial testing but negative on repeat single testing. However, all of the samples were negative for HCV RNA by RT-PCR method.
Conclusions
No sample was found to be serologically negative and HCV RNA positive. However, further studies are recommended for further clarification.
Key words : Blood donation, Donor screening, Hepatitis C virus, plasma, Pooled, PCR
R. Amini , A.a. Pourfathollah , M. Kamgooyan , Sh. Samiee ,
Volume 2, Issue 3 (3-2005)
Abstract
Abstract
Background and Objectives
In this study our aim was to determine HLA-Class I and II antigens freguencies of Hamedani ethnic group. In addition to demographic studies and disease association, it has wide application in bone marrow donor registeries.
Materials and Methods
In order to establish DNA-based HLA typing in central Laboratory of Iranian Blood Transfusion Organization, a comparison of serological and molecular (sequence specific primers “SSP”) methods for HLA-DRB has been performed. The study was descriptive and the population under study were selected out of the native people of Hamedan 100 healthy volunteer blood donors were chosen by questionnaire. 10ml heparinized and 3ml EDTA blood were collected from each selected donor. EDTA (PCR) samples were then frozen.
Results
N.I.H standard microlymphocytotoxicity and Nylon wool T and B cells separation was used for serological I and II typing. HLA-Class I plates were prepared from Iranian Blood Fractionation and Research Company and for Class II we used Biotest DR/DQ Typing Trays. PCR was done using “Roche high pure DNA extraction” Kit and HLA-DRBSSP (Biotest). The most and least frequent HLA-B antigens were B5 group (B51/B52) and B16 (38,39) respectively. Because of low resolution of HLA-DRB Kit, no significant difference was observed between serological and PCR methods. Although some blanks have been determined by PCR.
Conclusions
The HLA-DRB determination by PCR is mandatory for donor/recipient pairs (even sibling) for bone marrow transplantation for donors it should be done by high resolution kits.
Key words : HLA, Ethnic, DRB, PCR
M. Moghtadaei , G.h. Edrissian , S. Amini Kafiabad , Sh. Samiei , H. Keshavarz , M. Nateghpoor ,
Volume 2, Issue 4 (6-2005)
Abstract
Abstract
Background and Objectives
After hepatitis and AIDS, malaria is the most prevalent transfusion outcome in endemic areas. Presence of asymptomatic carriers of malaria parasites in the endemic areas can be a source of infection in transmission of malaria by blood transfusion. Prevention of malaria caused by blood transfusion depends on screening blood donors and deleting infected blood samples. To screen blood samples, parasitological, serologic and molecular methods have been applied.
Materials and Methods
In this study 120 blood donors in Iranshahr in Sistan-Baloochestan province were tested with different methods of thick and thin blood films, Immuno-Fluorescent Antibody Test (IFAT), and Polymerase Chain Reaction (PCR).
Results
The result of all thick and thin blood films were negative. IFAT by using P.vivax antigen and P.falciparum antigen for 38 and 6 donors respectively showed a titre of antibody equal to
± 1/20-1/320 (17 of the former group and 4 of the latter had a history of malaria infection). The PCR assay using silica for DNA extraction and using P.falciparum specified primers with sensitivity rate equal to 2-3 parasites per microlitre of blood was negative for all subjects under study.
Conclusions
This study showed, although microscopic examination of blood smears was inexpensive and simple, but it is labor-intensive and time-consuming that makes it insensitive for detection of low-level parasitemia in asymptomatic donors and for screening a large number of specimen. IFAT would not always show the real existence of parasites and in spite of simplicity and sensitivity because of its disability to be automated is not suitable for screening a large number of specimen. On the other hand, IFAT in individuals with malaria history and absence of parasites in their blood may be positive for a long period. It was approved that molecular methods such as PCR were more sensitive and more specific than conventional microscopic examination and their great advantage was the ability to detect the infection with low-level parasitemia that may have been distinguished by blood films examination. In the present study, probably because of low number of specimen or limited study duration with PCR method, or probably since parasitemia exiting in the subjects under study was less than 2-3 parasites per microlitre of blood, we were not able to detect positive cases.
Key words: Malaria, Blood transfusion, PCR, IFA, Blood donors
H. Ghaffari , K. Alimoghaddam , F. Foroughi , B. Chahardouli , Z. Sanaat , B. Bahar , A. Mousavi , M. Iravani , A. Ghavamzadeh ,
Volume 2, Issue 5 (9-2005)
Abstract
Abstract
Background and Objectives
The co-existence of recipient’s and donor’s hematopoietic systems after allogeneic marrow transplantation is called mixed chimerism. Chimerism analysis provides a national method of assessing the ability of different conditioning regimens, graft-versus-host disease (GVHD), prophylactic regimens, and cellular therapy to promote engraftment.
Materials and Methods
The association of mixed chimerism with acute graft-versus-host disease (GVHD), disease recurrence, survival, and relapse free survival was investigated in 91 patients 12 and 79 of whom underwent either bone or peripheral blood HLA-identical marrow transplantation respectively. Chimerism was assessed using multiplex amplification of shorty tandem repeats (STR-PCR). Cases included thalassemics (19 subjects), AML (29), ALL (20), CML (18) and others (5). Median age was 21 (age range of 3-50). There were 38 females (41.8%) and 53 males (58.2%). Conditioning was busulfan plus cyclophosphamide in 34 patients, busulfan plus fludarabin in 51 patients and busulfan plus fludarabin plus anti-thymocyte globulin in 6 patients. Median time of follow up was 13 months. Data was analyzed using SPSS statistical software.
Results
On day 30 after transplantation, mixed chimerism (MC) was observed in 15 patients (16.5%), complete donor chimerism (CC) in 72 patients (79%), and no chimerism in 4 patients. The incidence of acute GVHD was significantly lower in mixed chimeras than in complete chimeras (p=0.01) but there was no significant difference in acute GVHD grade (I, II vs. III, IV) between two groups. The incidence of relapse and overall survival were 17.6% and 88.9% respectively showing no significant difference between MC and CC. Relapse free survival was 80.2% and not significantly different between two groups.
Conclusions
Despite some previous reports, we found no significant difference in survival and relapse rate between MC and CC.
Key words: Allogenic, Bone Marrow Transplantation , Chimerism, PCR
Shaiegan M., Tarabadi F.a., Amini Kafi-Abad S., Samiei Sh., Babaeie Gh., Talebian A.,
Volume 2, Issue 6 (1-2006)
Abstract
Abstract
Background and Objectives
Beta-2 microglobulin ( β 2MG) is the light chain of Histocompatibility–Class I human antigen and its normal range is <3mg/ml. β 2MG level in sera of hepatitis B patients increases. In Hepatitis infection the presentation of the viral antigen on the hepatocyte in the presence of Class I HLA antigen plays a major role in the elimination of the virus.
Materials and Methods
In this descriptive study, s β 2MG, HBsAg (by ELISA), and HBV DNA (by PCR) were evaluated in sera of 49 patients with hepatitis B and 35 subjects in control group .
Results
Our results showed HbsAg was positive in all patients. 29 of patients were HBV-DNA-PCR positive and 20 HBV-DNA-PCR negative. β 2MG in all subjects in control group was in normal range and in 34.7% of patients above normal limit. β 2MG in HBV-DNA-PCR positive patients was higher than HBV DNA PCR negative patients. Such differences were significant (p < 0.05) .
Conclusions
It seems S β 2MG is a good marker for HBV replication and its absence may exclude HBV replication. The role of β 2MG in monitoring response to therapy needs to be further evaluated.
Key words: Hepatitis B virus, HBsAg , β 2MG (beta - 2 microglobulin), PCR , ELISA
F Rahmdani, S.a Mesbah-Namin, T Tarihi,
Volume 3, Issue 2 (6-2006)
Abstract
Abstract
Background and Objectives
RhD antigen has an important role in causing hemolytic diseases in newborns in testing fetal RhD status by an invasive method such as amniocentesis, it would also harm the fetus and mother. This study tried to detect fetal RhD genotype using free fetal DNA in maternal serum by a noninvasive method using hemi-nested PCR.
Materials and Methods
Blood samples were collected from 45 RhD- negative pregnant women whose spouses were Rh-positive the women's serum free DNAs were isolated by Phenol-Chloroform method. The exon 10 of the RhD genes was amplified in two rounds of PCR using special primers. For confirmation of negative results obtained in the second PCR, the fragment of the RhCE gene was amplified using special primers.
Results
Follow up and evaluation of the PCR results and relevant serological analysis of cord blood after delivery revealed that in 37 out of 41 cases the RhD had been determined appropriately (95.45%). There were one false-negative and three false-positive cases.
Conclusions
This study showed that maternal serum is very suitable for prenatal diagnosis of fetal RhD using noninvasive methods with minimum risk for neonates. In this regard, modern molecular methods with high sensitivity such as hemi-nested PCR and elimination of interfering factors could be applicable in clinical approaches.
Key words : Pregnancy , Serum, RhD, RhCE, PCR
Hemmaty T, Ghaffari H, Alimoghaddam K, Gharehbaghian A, Rostami Sh, Ashouri A,
Volume 3, Issue 3 (10-2006)
Abstract
Abstract
Background and Objectives
WT1 gene encodes a transcription factor that is involved in differentiation and proliferation of hematopoietic precursor cells as well as some other tissues like kidney, ovary, heart, etc. Quantitative assessment of WT1 gene expression is proposed as a useful marker in MRD detection and leukemia management.
Materials and Methods
To assess the relevance of this gene, we analysed peripheral blood mononuclear cells of 62 AML patients (new cases) for the expression level of WT1 mRNA using Real-time quantitative RT-PCR. We followed the analysis up to 3 years, depending on patient availability. This study as a fundamental and applicable one was done cross-sectionnaly. Samples were obtained randomly from the AML patients referred to the BMT center, and selection was based on the diagnostic criteria defined by clinical wards. WT1 expression in MNCs of patients was compared with 24 healthy individuals (K562 cells considered to express WT1 gene equivalent to 106).
Results
Samples for diagnosis showed significantly high levels of WT1 expression (>80%). After chemotherapy, its expression decreased (diminished about 1-2 log within induction therapy and around 3-4 log after consolidation therapy). There was a noticable correlation between the relative expression levels of WT1 and predicition of relapse (lower than gray zone versus higher than). Patients whose WT1 expression levels remained lower than the gray zone benefit from a better compete remission. On the contrary, 1-6 months prior to overt clinical relapse in 6 patients, their WT1 expression raised to levels upper than gray zone.
Conclusions
This study revealed that WT1 is a useful marker for detecting minimal residual disease, assessing chemotherapy effects, and prediciting relapse in AML patients.
Key words : WT1 gene, Minimal Residual Disease (MRD), PCR
Mohammad Reza Abbaszadegan, Massod Ziaee, Farhad Khadivi-Zand, Zahra Badiee, Bahram Khazaei, Zohreh Vahedian, Fahimeh Mehrabian, Ezat Dadkhah,
Volume 3, Issue 4 (1-2007)
Abstract
Abstract
Background and Objectives
Hemophilia A is the most common X-linked blood coagulation disorder. The prevalence rate of this disease in various communities is about 1-2/10000 males. The prevalence of hemophilia A is very high in southern Khorasan population, perhaps due to their isolation and high rate of consanguinity. The aim of this research was to detect hemophilia A carriers in two villages of southern Khorasan and set a prenatal diagnosis program for these families.
Materials and Methods
Blood samples of 34 patients with hemophilia A out of 51 family members (9 families) from two villages were collected. We were also able to collect samples from 7 mothers out of 9 families. Intragenic polymorphic sites in factor VIII gene including HindIII, Bc1I and AlwNI were used for carrier detection. DNA extraction and PCR-RELP were also performed.
Results
The results revealed 3 heterozygote mothers for HindIII, 2 for Bc1I and 1 for AlwNI. We utilized these polymorphic sites for carrier analysis in these families. Ultimately, 4 carrier sisters of affected boys in this population were found it was then possible to perform prenatal diagnosis procedure for their families.
Conclusions
HindIII and Bc1I polymorphisms can be suitable markers for hemophilia prenatal diagnosis in these two villages.
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Key words : Hemophilia A, RFLP (Restriction Fragment Length Polymorphism), Carrier detection�
Javad Douzandeh, Mehrdad Ravanshad, Saeed Amel Jamehdar, Farzaneh Sabahi,
Volume 3, Issue 4 (1-2007)
Abstract
Abstract
Background and Objectives
Human Immunodeficiency Virus type 1 (HIV-1) is the causative agent of Acquired Immunodeficiency Syndrome (AIDS) in man and diagnosis of HIV-1 genome in suspicious specimens is of special importance. Viral transmission through blood and blood products during the window period is considered to be an important concern. In addition, the employment of rapid, sensitive and accurate techniques is highly necessary for diagnosis of HIV-1 prior to antibody production in infants born from infected mothers. In the present study, a sensitive and rapid RT-Nested PCR to detect viral genome has developed.
Materials and Methods
In this study, a sensitive RT-Nested PCR technique for detection of a conserved HIV-1 "gag gene" sequence was developed. By using a specific primer pair, the fragment was amplified in two rounds of PCR. In order to confirm positive results, the developed technique was applied to standard Iranian panel as well as to positive specimens from different infection and disease categories in which reactivity was proven by confirmatory HIV-1 tests (ELISA and western blot).
Results
Thirty five sera from different stages of the disease as well as 15 standard Iranian panels and 20 negative control sera were collected and tested by the developed technique. In positive cases, a specific band was observed on agarose gel electrophoresis, while no band could be detected for negative control sera.
Conclusions
In this study, it was demonstrated that the developed HIV-1 RT-Nested PCR has a high sensitivity and specificity for diagnosis of HIV-1 infection. It has the advantage of viral genome detection prior to seroconversion and can be easily applied to detect HIV-1 infection during the window period.
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Key words : Nested PCR , RT-PCR, HIV-1, Gag gen�
S. Amini Kafi-Abad, A. Talebian, M. Moghtadaie, F. Ranjbar Kermani, F. Ferdowsian, Sh. Samie, A. Taghi Nia, M. Sobhani, Z. Ataie, M. Kavari,
Volume 3, Issue 5 (1-2007)
Abstract
Abstract
Background and Objectives
The incidence of post transfusion hepatitis has been reduced by blood donor screening for HBsAg, but the HBV infection is still responsible for certain cases of post-transfusion hepatitis world-wide. An estimate of the rate of HBV DNA and anti-HBc positive units is important for evaluation of the need for anti-HBc blood donor screening. In this study, the HBsAg negative blood units were evaluated for anti-HBc and all of anti-HBc positive units were tested for HBV DNA by PCR method.
Materials and Methods
Extra samples were collected from 2000 HBsAg, anti-HCV, anti-HIV and RPR-negative blood donors. All of the samples were examined by the approved anti-HBc assay. All anti-HBc positive samples were tested by anti-HBs assays and evaluated for HBV DNA (PCR). The sensitivity of the HBV DNA (PCR) assasy was estimated to be 300 geq/ml according to VQC proficiency panels.
Results
230 (11.5%) out of 2000 samples were positive for anti-HBc. 179 (77.8%) out of 230 anti-HBc positive samples were HBsAb positive, and 51(23.2%) HBsAb negative. All 230 samples were assayed for single HBV DNA (PCR) 227 of which came out to be negative for HBV DNA (PCR). Three blood donors were recalled and new samples from two of whom were collected. These new samples were negative for HBV DNA.
Conclusions
Further study for evaluation of HBV DNA in anti-HBc positive blood units with full automatic instruments and usage of blood bags with accessories is strongly recommended.
Key words: PCR, Blood donors, HBs Ag
M.a. Jalali Far, T. Zandieh, S.j. Imam, H. Galehdari, A.a. Pourfathollah, M. Baba Ahmadi,
Volume 3, Issue 5 (1-2007)
Abstract
Abstract
Background and Objectives
Occurrence of new infectious agents threatens access to zero risk in blood transfusion and enhancement of blood safety. Although sensitive methods are available for diagnosis of hepatitis, yet some hepatitis cases do not have a known etiology. In 1997, the novel DNA virus was isolated from post-transfusion serum samples of patients affected by non-A-G hepatitis. Nowadays this novel virus is known as transfusion-transmitted virus. This circular single stranded unenveloped and virucidally resistant virus is the first human circovirus and has universal distribution. It is believed that TTV may cause hepatitis and aplastic anemia. This study was conducted to determine the prevalence of TTV in healthy blood donors in Ahwaz and set up N22 PCR for subsequent first-time viral studies in south region in Iran.
Materials and Methods
In 2003, We studied the presence of TTV DNA by using Okamoto primers with PCR in plasma of blood donors in whom serologic tests for hepatitis A-C and HIV-Ab were negative.
Results
Our study showed that the virus prevalence in blood donors was 23.7% (60/253) and there was not any significant differences between prevalence of TTV and background variables.
Conclusions
Our findings showed the same prevalence rate as in neighboring countries however, in comparison with thalassemic patients that were studied in parallel with the present research, the difference was significant (143/250 57.3%). It shows the importance of blood transfuison in transmission of the virus.
Key words : Torque Teno Virus, Blood Donors, Iran, Blood transfusion, P
N. Obeidi, H. Ghaffari, A. Gharehbaghian, K. Alimoghaddam, M. Dehghan, A. Ghavamzadeh, A. Shamshiri,
Volume 4, Issue 1 (3-2007)
Abstract
Abstract
Background and Objectives
Cytomegalovirus (CMV) is an important pathogen in patients undergoing bone marrow transplantation. The prevalence of CMV varies from 30-100% in different countries as shown by seroepidemiological studies. Only 20-25% of patients develop CMV disease. Because of the similarity between CMV and GVHD and the different therapies required, detection of viral load will be effective in patients' survival.
Materials and Methods
51 recipients of BMT were monitored for 100 days post-BMT during which the samples were collected weekly. The Real-Time PCR was developed for quantitation of CMV viral DNA, using TaqMan tecnnology. For generation of standard curve, UL83 gene from CMV was cloned into a plasmid using a T/A cloning procedure. RQ-PCR assay was preformed in parallel with pp65 antigenemia assay on 415 samples.
Results
The results obtained by both techniques were significantly correlated (p<0.01). We could detect 13×101 -15×107 (CMV DNA copies/2×105 cells) by RQ-PCR method. About 76% of patients developed at least one episode of CMV reactivation. First positive result of RQ-PCR appeared 13 days earlier than of pp65 antigenemia. After preemptive therapy, 16 days were required to achieve negative result by RQ-PCR.
Conclusions
Both assays were highly correlated however, RQ-PCR was more sensitive than the antigenemia assay. After preemptive therapy, negative results of RQ-PCR were the best indicator to determine the endpoint of treatment and its success.
Key words : Cytomegalovirus (CMV), Bone Marrow Transplant (BMT), PCR.
Y. Mortazavi, M. Behzadi Fard, A.a. Pourfathollah, S. Kaviani, A.a. Feizi,
Volume 4, Issue 1 (3-2007)
Abstract
Abstract
Background and Objectives
Acute myeloid leukemia (AML) comprises a heterogenic group of malignant disorders involving cell maturation arrest at an undifferentiated stage in bone marrow. Activation of N-RAS proto-oncogene due to point mutations plays a major role in AML malignancy. Since there was no report on the frequency of N-RAS gene mutations in Iranian AML patients, therefore, we decided to determine its frequency and compare the results with age, sex and FAB subtypes.
Materials and Methods
In this descriptive study, 60 de novo AML patients from Tehran Shariati hospital, hematology-oncology and bone marrow transplantation center were screened for the mutations of N-RAS gene at codons 12, 13 and 61. DNA was extracted from peripheral blood samples before the start of chemotherapy. The above mentioned codons were amplified by PCR and analyzed by restriction endnuclease enzymes.
Results
We were able to detect mutations in 12 out of 60 (20%) patients. Most of the mutations were detected in men with an age over 40 years old. The frequency of mutations for codons 12, 13 and 61 were 15% ,11.6% and 5% respectively. Most of the mutations (33.3%) were found to happen in AML-M4 FAB subtype. We could not detect any mutation in AML-MO, M6 and M7.
Conclusions
We detected mutations in 20% of our AML patients. In general, the frequency of the mutations we found was in agreement with the results of other studies. However, a study with more patients and a wider range of age using a combination of PCR-RFLP and direct gene sequencing is highly recommended.
Key words : Acute myelocytic leukemia, Mutation. PCR
T. Madani, Sh. Samiee, Z. Attaei, M. Kavari, Gh.r. Babaei, M. Mostakhdemin, M. Shaiegan,
Volume 4, Issue 3 (9-2007)
Abstract
Abstract
Background and Objectives
Serologic methodes used in HPA–typing are limited due to restricted access to specific anti-sera and decreased platelet count in thrombocytopenic patients. Therefore, several DNA-based HPA-genotyping techniques were used to determine the genotype of HPAs. Since nothing is known about the HPA gene frequency in Iran, this study was performed to determine its frequency in some Iranian blood donors.
Materials and Methods
DNA was extracted from a 3-ml whole blood sample prepared from donations of 100 Iranian blood donors collected in EDTA-coated blood tubes. Human platelet (PLT) alloantigens (HPA)-1/2/3/4/5 and HPA-15 typing were performed by the Polymerase Chain Reaction – Sequence Specific Primer technique ( PCR-SSP), and HPA-1a phenotyping was performed by ELISA method for 40 % of samples.
Results
The frequencies of HPA genes were: HPA-1a 98% , HPA-1b 2% , HPA-2a 54% , HPA-2b 46% , HPA-3a 48% , HPA-3b 52% , HPA-4a 100% , HPA-5a 99% , HPA-5b 1% , HPA-15a 47% , and HPA-15b 53%. HPA-4b was not found . The frequencies of HPA phenotypes were determined to be: HPA1a/1a 96% , HPA1a/1b 4% , HPA2a/2a 8% , HPA2a/2b 92% , HPA3a/3a 19% , HPA3a/3b 59% , HPA3b/3b 22%, HPA4a/4a 100% , HPA5a/5a 98% , HPA5b/5b 2% , HPA15a/15a 14% , HPA15a/15b 67% , and HPA15b/15b 19%. 40 HPA-1a phenotyping by ELISA showed 26 positive (OD > 0.5 ), 4 negative (OD < 0.3 ), and 10 indeterminate samples (0.3 < OD < 0.5) .
Conclusions
No HPA-1b/b homozygous genotype similar to other Asian studies was found. Since HPA-1,-2,-5 frequencies in the population under study differ from the European Caucasian race, it seems that antibody production in our population might be different from other Caucasians. According to HPA frequencies, it seems that HPA-2b, HPA-5b and HPA-15b may induce posttransfusion purpura and platelet refractoriness which need further investigation.
Key words: Platelet antigens, Gene frequency, PCR, Iran
F. Yari, N. Bagheri, M. Zaman Vaziri, M. Sobhani, F. Sabaghi, A. Talebian,
Volume 4, Issue 3 (9-2007)
Abstract
Abstract
Background and Objectives
Major Histocompatibility Complex (MHC) is the most polymorphic system in the genome of different species. In human beings, these genes named HLA are located on the chromosome 6. HLA class I and II undertake genetic control of the immune system. Identification of HLA alleles is useful in transplantation, disease, and anthropological studies. Among different antigens of HLA, DR (DRB1) antigens are the most variable. In this research, due to the importance of DRB1 antigens in bone marrow transplantation, these antigens were studied in normal population. This study was performed on different Iranian races and did not suffice to a specific group.
Materials and Methods
DNA was extracted from the whole blood sample of 466 normal individuals after randomized sampling. Some HLA-DRB locus segments were amplified using 23 primer pairs by using PCR-SSP method. Finaly, PCR products were evaluated by electrophoresis in 2% agarose gel.
Results
The most prevalent alleles in DRB1 locus in normal poulation of Iran were DRB1*11, DRB1*13, DRB1*15, and DRB1*04 (20%, 11.4%, 11.4%, 10%, respectively). Whereas DRB1* 09 was the least frequent allele.
Conclusions
This research showed genetic diversity of HLA DRB1 in the mixed Iranian population. The data suggest that the Iranian population share certain HLA class II genetic components with the populations residing in Russia and Eastern and Southern European countries.
Key words : HLA-DRB1, PCR, Iran
A. Jafari Kermani, F. Fathi, S.j. Mowla, Y. Gheisari,
Volume 4, Issue 4 (2-2008)
Abstract
Abstract
Background and Objectives
Stem cells are defined with two main characteristics which are self renewal potential and pluripotency. It is of great importance to study expression of self renewal genes in different stem cells, because self renewal regulatory mechanisms may vary among different kinds of stem cell. Human umbilical cord stem cells have recently gained so much attention, because their use in cell therapy methods has several advantages. The main objective of the present study was to evaluate the expression of Oct-4, Sox-2, Nanog, Nucleostemin, Bmi-1 and Sox-9 self renewal genes in human umbilical cord vein mesenchymal stem cells.
Materials and Methods
In this experimental study, umbilical cords were obtained in sterile condition and carried to the laboratory in HBSS buffer. Mesenchymal stem cells were harvested by collagenase treatment and cultured by means of their adhesiveness to plastic surfaces in DMEM medium supplemented with 15% FBS. To confirm the mesenchymal identity of these cells, expression of some surface markers including CD105, CD106, CD54, CD45, HLA-DR, CD166, and CD34 was analysed by means of flowcytometery. Finally, expression of self renewal genes was evaluated by RT-PCR and immunocytochemistry techniques.
Results
Flowcytometry analysis of UBMCs revealed that the cells are positive for CD105 (98%) and negative for CD34 (0%), CD54 (1%), CD45 (0%), CD106 (0%), CD166 (0%), and HLA-DR (0%). Our results also revealed Oct-4, Nanog, Nucleostemin, Bmi-1 and Sox-9 expression in human umbilical cord vein mesenchymal stem cells, but not Sox-2 self renewal gene expression.
Conclusions
The results showed that the mechanism which is known today for Oct-4, Nanog and Sox-2 function is not the only possible mechanism in which these three genes can play role to regulate self renewal potential of stem cells. Obtained results can offer that a combination of regulatory mechanisms which regulate self renewal of adult and embryonic stem cells, may be responsible to regulate self renewal of stem cells.
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Key words : Mesenchymal stem cells, Umbilical cord, PCR, Immunocytochemistry
F. Mahmudi, Dr. M. Mahmoodian Shooshtari, Dr. Z. Sharifi, Dr. M. Hosseinni,
Volume 5, Issue 1 (3-2008)
Abstract
Abstract
Background and Objectives
Human parvovirus B19, a member of the parvoviridae family, with single-stranded DNA is a very minute and non-enveloped virus. Human parvovirus B19 causes a number of clinical illnesses including infectious erythema (fifth disease), hydropes fetalis, transient aplastic crises, arthropathy and congenital aplasia. B19 virus is transmitted via respiratory tract, blood products, and blood transfusion. Parvovirus B19 DNA is detectable through molecular techniques such as PCR. Previous studies indicate that individuals who have anti B19 IgG are negative. IgM antibody against B19 lasts for a few months and IgG antibody persists for many years or lifelong. The aim of this study was to evaluate the prevalence of B19 among blood donors in Tehran.
Materials and Methods
In this cross-sectional study, sera of 730 blood donors who were negative for HIV, HBsAg and HCV were tested for IgG and IgM anti-B19V using ELISA. Then, all of the sera were tested for presence of B19 DNA through semi-nested PCR.
Results
Out of 730 blood donors, 4 (0.5%) had IgM antibody thereby being reported positive 338 subjects (46.3%) were positive for anti-B19 IgG. DNA B19 was not found in any of the subjects (0%).
Conclusions
Although B19 may cause adverse reactions in some blood recipients, screening blood donors for B19 is not essential. In this study, the high prevalence of IgG on one hand and the lack of any positive cases of B19 DNA on the other indicate of the low risk of the latter to be transfusion transmittable. Further studies on blood donors particularly high risk groups are recommended.
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Key words : Parvovirus, Blood donors, Prevalence, PCR
M.h. Moghaddasi, Sh. Samiee, S. Amini Kafi Abad, Z. Attaee, M. Kavari, M. Sobhani,
Volume 5, Issue 2 (8-2008)
Abstract
Abstract
Background and Objectives
Thrombophilia is a common and dangerous disease. It may be heritable or acquired. A substitution in the 3' untranslated region of prothrombin gene (PRT-G20210A) has been reported to be related to thrombophilia in Caucasians, but this relationship remains in debate in other populations. The hetrozygote form of PRT G20210A gene variant expresses prothrombin 25% higher than the average level. This study was performed to evaluate the prevalence of G20210A gene mutationas in thrombotic patients.
Materials and Methods
In this descriptive and cross sectional study, 299 patients with venous thromboembolism were selected. The subjects consisted of 116 male and 183 female. Traditional risk factors for venous thrombosis and pulmonary thromboembolism such as proteins C, S, ATIII deficiency and APC-resistance were investigated as well.
Results
We found that only 1.7% of patients with venous thromboembolism were carrier of prothrombin G20210A mutation which is one of the prevalent point mutations in Caucasians patients (18%). The homozygote form of PRT G20210A was not found.
Conclusions
Prothrombin G20210A mutations in Iranian population as opposed to western populations are rare (1.7%) with APC-resistance being common in the former.
Key words: Prothrombin, Mutation, PCR, Venous thromboembolism
S.h. Ghaffari, M. Yaghmaie, K. Alimoghaddam, A. Ghavamzadeh, M. Jahani, S.a. Mousavi, M. Irvani, B. Bahar, E. Baibordi,
Volume 5, Issue 2 (8-2008)
Abstract
Abstract
Background and Objectives
A specific chromosomal abnormality, the Philadelphia chromosome (Ph), is present in more than 90% of CML patients. The aberration results from a reciprocal translocation between chromosome 9 and 22, creating a BCR-ABL fusion gene. Major forms of BCR-ABL fusion are b3a2, b2a2, and e1a2. Less common fusion genes are b3a3 or b2a3 (p203) and el9a2 (p230). The aim of this study was to set up a multiplex RT-PCR assay and determine the frequency of different fusion genes in 75 Iranian patients with CML.
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Materials and Methods
In this experimental study, peripheral blood samples from 75 adult Iranian CML patients were analyzed by multiplex RT-PCR to detect different types of BCR-ABL transcripts of the t(9:22).
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Results
All of the examined cases were positive for some type of BCR/ABL rearrangement. The majority of patients (82.6%) expressed one of the P210 BCR-ABL transcripts (b3a2, 62% and b2a2, 20%), while the remaining showed either one of the transcripts of b3a3, b2a3, ela2 or coexpression of b3a2 and b2a2. The rate of coexpression of the b3a2 and b2a2 was 5%.
Conclusions
It is possible to detect rare fusions other than common ones through multiplex method. In Iranian CML patients, b3a2 was three times more frequent than b2a2 shown to be higher as compared with other studies. Probably, the kind of fusion is of clinical importance and helps us in understanding t(9:22) leukemic cells pathogenesis.
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Key words : Chronic myeloid leukemia, RT-PCR, Philadelphia chromosome
Dr. M. Bouzari, A. Mokhtarzadeh,
Volume 5, Issue 4 (1-2009)
Abstract
Abstract
Background and Objectives
In 1997, a novel DNA virus, unrelated to the known hepatitis viruses, was isolated from the serum of a patient with posttransfusion acute hepatitis with elevated alanine aminotransferase of unknown etiology, and it was named TT virus (TTV). In this study, the frequency of TTV and its role in induction of liver damages were evaluated in three groups of healthy, HBV and HCV-infected blood donors in Tabriz.
Materials and Methods
In this descriptive study, sampling was done on 100 healthy donors, 40 HBsAg positive and 40 positive Anti-HCV in East Azarbayjan Blood Transfusion Center. The levels of ALT and AST in the sera were measured DNA was then extracted and PCR was performed using T801 and T935 consensus primers. One-way ANOVA, Tukey-Kramer, Kruskal-Wallis, Dunn's and Fisher’s Exact tests were finally used for statistical analyses.
Results
TTV was detected in 65%, 70% and 67.5% of the healthy, HBV and HCV-infected blood donors, respectively. In each group of blood donors, the levels of ALT and AST were not significantly different in TTV infected versus non-infected individuals.
Conclusions
Given the high frequency of infection in healthy individuals and considering the level of hepatic enzymes in TTV infected individuals, it seems that the virus or at least certain genotypes present have not been pathogenic for the infected individuals tested.
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Key words : Torque teno virus, Blood donors, Alanine Amniotransferase, Polymerase Chain Reaction
