Showing 7 results for Rt-Pcr
R Arvan , E Elahy ,
Volume 56, Issue 5 (7-1998)
Abstract
Macrophage colony stimulating factor (M-CSF) has previously been shown to affect the differentiation of cells of the mono-nuclear phagocytic line. More recent studies indicate that M-CSF may have a role in pregnancy. In the present study, the expression of M-CSF in the human placenta was demonstrated. Placental mRNA was isolated and used as template for synthesis of complementary DNA (cDNA). The presence of M-CSF related sequences in the cDNA was shown by PCR and RT-PCR reactions in which M-CSF specific primers were used. In addition, it was shown that a 2.4 kb cDNA after electrophoresis and transfer to a nylon filter, hybridized with a digoxygenin labeled M-CSF specific probe.
Ahani R, Derakhshandeh Peykar P, Raoofian R, Heidari M,
Volume 67, Issue 1 (4-2009)
Abstract
Background: Leukemia is one of the most common pediatric malignancies. T-cell Acute Lymphoblastic Leukemia (T-ALL) accounts for 15% of hematopoetic cancers. It has been well understood that identification of genetic alterations associated with leukemias is very critical. The molecular genetic techniques have promoted the identification of leukemia-associated genetic changes that may characterize the most accurate predictors of clinical outcome. These considerations reinforce the requirement for rapid identification of the abnormalities.
Methods: Multiplex RT-PCR, a highly sensitive and specific method applied to screen simultaneously three most frequent transcription factors, TLX1/HOX11, TLX3/HOX11L2 and TAL1/SCL which are associated with T-cell Acute Lymphoblastic Leukemia (T-ALL).
Results: We describe here our efforts to establish a multiplex RT-PCR analysis system that facilitates the detection of HPB-ALL and K562 cell lines, respectively.
Conclusion: The multiplex RT-PCR technique is a sensitive, valuable and cost-effective diagnostic tool which could improve our ability to accurately and rapidly risk-stratification of patients with childhood T-ALL. In order to perform multiplex RT-PCR technique researchers do not need bone marrow samples and they can employ this method using peripheral blood samples. Therefore, the status of treatment could be followed by assessment of the level of mRNA expression of oncogenic transcriptional factor using peripheral blood sample. Use of this procedure not only provides the best results in short term for specialist, but also clinicians could have opportunities to choose suitable treatment strategies with decrement of drug side effects.
Yahyazadeh Sr, Mehraban D, Ghaffari Sh, Alimoghadam K, Ghavamzadeh A, Naderi Gh, Kazemeyni Sm, Rasteh M,
Volume 67, Issue 1 (4-2009)
Abstract
Background: Transitional Cell Carcinoma (TCC) of bladder is the second most common urogenital malignancy and because of its high rate of recurrence (two third of tumors recur) vigilant surveillance is necessary. There have been a lot of efforts to find a proper biomarker for detecting urothelial cancers because available methods are expensive and invasive (like cystoscopy) or have a low degree of sensitivity (like urine cytology). Urothelial malignancies, like other cancers tend to express a large amount of telomerase. The aim of this study was to evaluate the possible application of voided urine human telomerase reverse transcriptase (hTERT) mRNA assay in detecting low-grade bladder carcinoma in comparison with urine cytology.
Methods: Voided urine samples were collected from 49 patients who were supposed to go under operation. Samples were examined by both Quantitative Real-time RT-PCR (for measuring hTERT mRNA level) and cytology the results were then compared to the final pathologic studies.
Results: Regardless of clinical stage and or pathological grade of tumor, sensitivity of telomerase test and urine cytology was 74% and 16% respectively. There was a strong correlation between results of urine cytology and stage and/or grade of tumor however, sensitivity of telomerase test was acceptable regardless of stage and or grade of tumor. There was a statistically significant difference between sensitivity of urine cytology and telomerase test (p<0.001).
Conclusion: Detection of hTERT-mRNA can potentially be used as a non-invasive method for diagnosis and follow up of bladder carcinoma instead of urine cytology.
Rahbarimanesh A, Saberi H, Modarres Gilani Sh, Salamati P, Akhtarkhavari H, Haghshenas Z,
Volume 69, Issue 8 (11-2011)
Abstract
Background: Acute gastroenteritis is a major cause of
morbidity and mortality among children in developing countries.
Rotaviruses are recognized as the most common etiologic factors of
gastroenteritis. In this study, we determined the epidemiologic
features, clinical symptoms and molecular structure of rotavirus VP4(P)
genotypes in children with acute diarrhea in Bahrami Hospital in Tehran
Iran, during 2009 for justifying the routine use of rotavirus vaccines
in children.
Methods: One hundred fifty fecal samples from
150 children with acute diarrhea in Bahrami Pediatric Hospital in
Tehran, Iran were collected from January to December 2009. The patients’
mean age was 20.90+18.19 years (ranging from 1 month to 14 years).
Fecal samples were transported on ice to the laboratory of virology
department of Pasture Institute of Iran. The demographic and clinical
data for each case were entered in an author-devised questionnaire.
Group A rotavirus was detected by dsRNA-PAGE. Subsequently, rotavirus
genotyping (VP4) was performed by semi-nested multiple RT-PCR and the
phylogenetic tree of the Rotavirus nucleotides was constructed. The data
were analyzed by statistical tests including Wilcoxon signed and
Mann-Whitney U.
Results: Rotavirus was isolated in 19.3% of the
samples, more than 90% of which had long RNA patterns. The predominant
genotype (VP4) was P[8] (86%) and other genotypes respectively were P[6]
(6.9%) and P[4] (6.9%).
Conclusion: A high prevalence of the P[8]
genotype was found to be the cause of acute diarrhea. The analysis of
P[8] genotype sequence showed a high level of similarity of the virus in
this study with those of other Asian countries.
Farhadi M, Tabatabaee A, Shekarabi M, Noorbaksh S, Khatib M, Javadinia Sh,
Volume 69, Issue 9 (12-2011)
Abstract
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Background: Too many studies are in the process
of determining the probable role of immune system in the etiopathogenesis of
nasal polyposis. This study was designed to identify the probable participation
of Th1, Th2 lymphocytes in the induction and progression
of nasal polyposis.
Methods: Seventy-five patients, 42 male and 33 female, with
nasal polyposis were examined for total serum IgE, specific serum IgE and reaction to skin test for differentiating allergic from
non-allergic participants in Rasoul Akram Hospital during 2010. To determine the possible correlation of allergic
reactions in the upper respiratory tract and nasal polyposis, cytokine gene
expression was evaluated on the extracted RNA by
RT-PCR. The data were analyzed by using c2, independent t-test, correlation and
Receiver operating characteristic (ROC)
curve.
Results: The mean
age of participants was 38 years (18-81 years). IFN-γ
and IL-4 gene expressions were more prevalent
in allergic than non-allergic individuals (IFN-γ:
39.5% vs. 14.2%, P=0.3 and IL-4: 44.7% vs. 18.9%, P=0.02, respectively). IL-10 and IL-12 (P35 and P40 fractions) genes were not
significantly different between the two groups. IL-10 and IL-12 (P35, P40) genes did not differ significantly either.
Conclusion: This research suggests that overproduction of
cytokines and an imbalance of Th1
and Th2 cell production may
play an important role in the pathophysiology of allergic or non-allergic nasal
polyp formation. Thus, although nasal polyposis is a multifactorial disease
with several different etiological factors, chronic persistent inflammation is
undoubtedly a major factor irrespective of the etiology.
Farhadi M, Tabatabaee A, Shekarabi M, Noorbakhsh S, Javadi Nia Sh, Ghavami Gh,
Volume 70, Issue 7 (10-2012)
Abstract
Background: Staphylococcus aureus secretes numerous superantigenes which trigger the inflammatory mechanisms of sinus mucosa and cause chronic rhino-sinusitis. This study was designed to evaluate the role of staphylococcus aureus superantigens in polyp tissues of patients with chronic rhino-sinusitis in comparison with a control group.
Methods: Polyp tissue samples of 28 patients and mucosal specimens of 19 healthy individuals were evaluated for staphylococcus aureus bacterium superantigens, exotoxins A, B, C and D and TSST-1 with RT-PCR and ELISA methods Rasoul Akram Hospital during 2 years.
Results: Polymerase chain reaction (PCR) results revealed that 88.2% of the patients and 45.5% of the controls had at least one type of superantigen (P=0.03). Evaluation of superantigens using ELISA method showed presence of at least one type of superantigen in the nasal samples of all patients and in 35.3% of the controls (P<0.001).
Conclusion: A relationship between staphylococcal superantigens and nasal polyps is concluded from this study which indicates the probable role of these superantigens in the pathogenesis of nasal polyposis.
Sh Savad, P Mehdipor, H Shirdast, L Nekoohesh, L Nekoohesh, R Shirkoohi, V Nikoui, M Miryounesi, Mh Modaresi,
Volume 71, Issue 2 (5-2013)
Abstract
Background: Breast cancer is the most common cancer in women. Non-coding RNAs especially miRNAs have important regulatory roles in cancer. MiRNAs are 21-24 nucleotides which have different levels of expression between tumors and normal tissues. In this study, we have analyzed expression level of miR-520d in three different groups of breast cancer.
Methods: Fifty nine samples were divided into different groups according to their immunohistochemistry (IHC) classification: estrogen receptor (ER) positive and/or progesterone receptor (PR) positive group (as group I) human epidermal growth factor receptor 2 (HER2) positive group (as group II) and Triple negative group (as group III). After small RNA extraction from tissues, cDNAs were synthesized and Real time RT-PCR carried out using DNA binding dye. Expression levels were analyzed by LinRegPCR and REST software.
Results: MiR-520d under- expressed in all of three different groups. The expression ratio in groups I ,II, and III were 0.193, 0.167, 0.21, respectively, but only the result from group II was significant (P=0.017). According to the different clinicopathological status of breast cancer, miR-520d underexpressed significantly not only in patients with metastatic lymph node (P=0.019) but also in patients which have cancer at stage III (P=0.036).
Conclusion: In this study, we found that miR-520d possibly acts as a tumor suppressor. It may be useful for diagnosis of tumor from normal tissue. In addition, miR-520d significantly underexpressed in HER-2 positive group of breast cancers. Therefore, it may be useful as an additional diagnostic test in this group of breast tumors along with other biomarkers.
