Showing 8 results for Pcr
Volume 2, Issue 1 (10-2009)
Abstract
Background: In 1997, a novel DNA virus was isolated from the serum of a patient in
Japan, and it was named TT virus (TTV). As the virus is replicated in liver and has the ability to induce apoptosis in hepatocytes (Hepatocellular carcinoma cells) it is hypothesized that TTV is an opportunistic virus and in certain conditions causes liver damage. In this study the frequency of infection with TTV was detected in two groups of healthy and hepatitis B infected blood donors in the
West
Azerbaijan
Province.
Material and Methods: Serum samples were collected from 100 healthy and 40 HBs Ag positive donors in west Azerbaijan Blood Transfusion Center. Patients’ characteristics, sex, age and blood groups were recorded. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the sera were measured and then DNA was extracted and polymerase chain reaction (PCR) was performed using T801 and T935 consensus primers to amplify a 199 bp segment of a much conserved non-coding region of the genome (UTR).
Results: TTV was detected in 69% and 75% of healthy and hepatitis B infected blood donors respectively. No significant difference was observed in the frequency of the infection with TTV in different blood groups or age groups (P>0.05). In each group of blood donors the level of ALT and AST were not significantly different in TTV infected and non-infected individuals (P>0.05).
Conclusion: Regarding 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 its investigated genotypes have not been pathogenic for the infected individuals examined.
Alireza Ghazimorad, Majid Bouzari, Mohammad Taghi Kardi,
Volume 6, Issue 3 (5-2014)
Abstract
Background: Recently, some new viruses have been identified for their association with hepatitis which Torque Teno Mini Virus being among them. The aim of this study was to determine the frequency of Torque Teno Mini Virus in healthy individuals and hepatitis B and C patients in Isfahan, Iran.
Materials and Methods: One hundred serum samples of healthy individuals from Isfahan Blood Transfusion Organization were collected. A total of 25 human serum samples from hepatitis B and 25 samples from hepatitis C infected patients were also collected from Mahdieh diagnostic laboratory in Isfahan, Iran. Viral DNA was extracted and Torque Teno Mini Virus DNA was detected using a nested PCR with primer sets designed for a conserved region of the Torque Teno Mini Virus genome. PCR and Reverse transcriptase PCR were used for detection of HBV and HCV respectively.
Results: Torque Teno Mini Virus -DNA was detected in 17% of healthy individuals. It also was detected in 20% and 48% of serum samples from hepatitis B and C infected individuals, respectively. The frequency of Torque Teno Mini Virus was significantly higher in hepatitis C patients versus healthy individuals (P < 0.05). Also, the frequency of TTMV in hepatitis C patients was significantly higher than hepatitis B patients (P < 0.05).
Conclusions: The difference in Torque Teno Mini Virus frequency between the hepatitis C and healthy group was significant (P< 0.05). The etiology of the higher infection rate in hepatitis C individuals needs to be determined.
Keywords: Torque Teno Mini Virus, Hepatitis B, Hepatitis C, PCR.
Pazhouhnia S, Bouzari M, Rahimi F, Kardi Mt,
Volume 7, Issue 1 (11-2014)
Abstract
Background: Approximately 600,000 deaths occur every year as a result of the acute and chronic consequences of hepatitis B virus infection. Ten different hepatitis B virus genotypes have been identified with distinct geographical distributions. Different clinical outcomes, including the rate of mutations, development of hepatocellular carcinoma, chronicity, response to treatment, transplantation rejection and occult infections, are affected by specific genotypes. The aim of the present study was to determine the frequency of genotype D of the virus in Isfahan, Iran.
Patients and Methods: In this study primarily hepatitis B virus positive patients were identified by the detection of HBs antigen using ELISA test and then PCR was used as a confirmatory test. Fifty five patients that were identified as hepatitis B positive were tested for hepatitis D genotype using type - specific PCR.
Results: The patients included 30 (54.5%) females and 25 (45.5%) males. In total, frequency of genotype D was 29 out of 55 cases (52.7%). Genotype D was detected in 19 (63.3%) females and 10 (40.0%) males indicating no statistically significant difference. The difference in the level of liver enzymes in patients infected with genotype D and non-genotype D hepatitis B virus were not significant.
Conclusion: In the present study the frequency of genotype D among patients with hepatitis B virus infection in Isfahan, Iran, was 52.7%. No significant relation was observed between the level of liver enzymes and infection with the genotype D.
Keywords: Hepatitis B virus, genotype D, PCR, Isfahan, Iran.
Morteza Bagheri, Isa Abdi Rad, Davood Maleki, Ali Eishi, Nasim Valizadeh,
Volume 10, Issue 2 (6-2018)
Abstract
Background: The Philadelphia chromosome (Ph) characterized by t (9; 22) (q34; q11.2) is a reciprocal translocation giving rise to a chimeric BCR-ABL fusion gene. Incidence of Ph chromosome is over 98% in Patients with Chronic Myeloid Leukemia (CML) and around 20% in acute lymphoblastic leukemia (ALL). The finding of this fusion gene is essential for diagnosis of CML by detection of various fusion transcripts such as b2a2 and b3a2 transcripts and Ph positive ALL by detection of e1a2 (p190) transcripts. We conducted this study to determine the frequency of various BCR-ABL fusion transcripts in the west Azerbaijani patients with CML.
Methods: RNA was isolated from peripheral blood samples by standard protocols. BCR-ABL fusion gene detection was carried out with one-step multiplex RT-PCR in 41 west Azerbaijani patients with CML.
Results: Among patients with CML, the frequencies of b2a2 and b3a2 transcripts were 52.5% and 12.5%, respectively. Co-expression of b3a2 and b2a2 transcripts was found in 12.5% of the patients.
Conclusion: The findings of this study showed that multiplex RT-PCR is a suitable technique to identify the typical BCR-ABL fusion transcripts in the west Azerbaijani patients with CML. Atypical transcripts possibly run away while using multiplex PCR.
Mohsen Hamidpour, Mohammad Ghorbani, Mostafa Rezaei-Tavirani, Hamid Reza Niazkar, Mohammad Reza Managhchi, Masoud Shahrudian,
Volume 11, Issue 3 (9-2019)
Abstract
Background: Factor V Leiden, Prothrombin gene (G20210A) and MTHFR (C677T) polymorphism are the main biomarkers for evaluation of tendency for venous thromboembolism. We aimed to investigate the frequency of mutations in factor V Leiden, Prothrombin G20210A and MTHFR C677T and identify the genetic status for these mutations in patients with venous thrombosis.
Methods: This study was carried out in 312 patients with venous thrombosis who were referred to “Thrombosis Clinical center”, Imam Khomeini Hospital, Tehran, Iran and “Sarvar Clinic”, Mashhad, Iran. Identification of gene mutations was performed using PCR-restriction fragment length polymorphism (RFLP)-based method.
Results: The prevalence of Factor V Leiden mutation was 35.8%, while 8.9% of them were homozygous for AA allele and 26.9% had the GA allele in heterozygous state. The prevalence of MTHFR (C677T) mutation was 17.9% of which 7.1% had the TT mutant allele in homozygous and 10.8% had CT allele in heterozygous state. The prevalence of mutation in prothrombin gene G20210A was 8.9% with all cases heterozygous for GA mutant allele.
Conclusion: In our study from two referral centers for thrombotic disorders, the prevalence of mutations in gene encoding factor V Leiden was higher than Prothrombin 20210A and MTHFR C677T polymorphisms. Therefore, assay for factor V Leiden mutation has the first priority in the evaluation of patients with hereditary thrombophilia in these geographical regions.
Dina El Dahshan, Amira Hammam, Aya Ahmed, Mohamed El Samra,
Volume 12, Issue 1 (3-2020)
Abstract
ackground: Despite extensive research in leukemia, intracellular events leading to prolongation of cell cycle and resistance to pro-apoptotic factors are still not clearly defined. In recent years, the search for such events led to focusing on an anti-apoptotic factor, PIM-2 (Proviral integration of Moloney virus-2). The aim of the present study was to assess the expression of PIM2 gene in patients with acute myeloid leukemia (AML) through quantitative real time polymerase chain reaction (QRT-PCR) and to correlate the results with clinical and laboratory findings of the patients as well as their response to the treatment. Methods: 80 patients with AML and control group were enrolled in the present study. QRT-PCR was used to study PIM2 gene expression.
Results: The mean expression level of PIM2 gene was significantly higher in AML patients (3.5941±7.7736) compared with the control group (0.5303±0.4014) (P=0.034). Its expression level was not different in terms of achieving remission. A positive correlation was observed between PIM2 gene expression and total leucocytic count (R=0.059, P=0.719), while there was a negative correlation between the gene expression and platelet count (R=-0.118, P=0.470). No significant correlation was found between PIM2 gene and patients’ response to treatment as (P=0.883) although it’s level was higher in patients who did not achieve complete remission (6.1±11.9) than patients who achieved complete remission (3.1±5.3).
Conclusion: The present study showed higher PIM2 expression level in AML patients than normal population. Also, its level was higher in patients who did not achieve complete remission.
Ghaida Raheem Lateef Al-Awsi, Shireen Ali Obaid, Saade Abdalkareem Jasim, Fadyia Mahdi Alameedy,
Volume 13, Issue 2 (6-2021)
Abstract
Background: Epstein-Barr virus (EBV)-associated post-transplant lymphoproliferative disorder (EBV-PTLD) is a known and serious condition for the hematopoietic stem cell transplantation. This study evaluated the prevalence of EBV in umbilical cord samples of healthy pregnant women in Baghdad, Iraq.
Methods: 800 umbilical cord blood (UCB) samples were collected from healthy pregnant women. The enzyme linked immunosorbent assay (ELISA) test was conducted to detect IgM and IgG antibodies. The EBV deoxyribonucleic acid (DNA) was detected in the plasma and buffy coat contents using quantitative real-time polymerase chain reaction (RT-qPCR) and nested PCR techniques.
Results: Four (IgG-positive), five and six samples were positive in the ELISA, Nested PCR and RT-qPCR assays, respectively. The age of contaminated women included 26-34 years. Notably, in the ELISA method, all the plasma samples were IgM-negative.
Conclusion: The existence of EBV among healthy women is a concern and assessment of a larger sample size is necessary to determine large-scale presence of EBV infection. The RT-qPCR exhibited higher sensitivity than ELISA and nested PCR techniques which can be considered as a confirmatory test. For larger sample size investigation, the ELISA method is appropriate as a primary screening test for lower costs.
Dr Sharareh Kamfar, Dr Fariba Alaei, Dr Reza Zaferani, Dr Vahide Zeinali,
Volume 14, Issue 4 (12-2022)
Abstract
Colorectal cancer (CRC) is the third leading cause of cancer-related death worldwide, and its occurrence can be ascribed to genetic susceptibility. Mitochondrial DNA 4977-bp (mtDNA 4977), as the most described mtDNA deletion, has been long proposed to be involved in various types of cancers. However, a few studies on mtDNA 4977-bp deletion in Iranian patients with CRC have been reported. The current study aimed to determine mtDNA 4977 frequency in CRC and its association with cancer susceptibility. We conducted a case-control study in which a total of 26 patients with CRC, 26 tumor tissues, adjacent normal tissues, peripheral blood samples, and peripheral blood samples from 50 healthy subjects were included. mtDNA 4977 was detected using multiplex PCR technique and direct DNA sequencing. Real-time PCR was also used to determine deletion levels. mtDNA 4977 was observed in six patients (23.07%), four (15.3%) in both tumor and matched surrounding normal tissues, and two (7.69%) in adjacent normal tissues, but not detected in both patients and control samples in peripheral samples. A significant difference was found between mtDNA 4977 deletion in tumoral and adjacent normal tissues (P=0.001). No relation was observed between mtDNA 4977 and categorical variables, including age and gender, and tumor stage. The analysis confirmed no association between the mtDNA 4977-bp deletion and susceptibility to colorectal cancer in Iranian patients. However, more extensive studies are required to confirm or reject these findings.
