Showing 6 results for Real-Time Pcr
Mahdi Paryan, Mahdieh Mondanizadeh, Samira Mohammadi-Yeganeh , Behzad Khansarinejad,
Volume 14, Issue 5 (11-2011)
Abstract
Background: HIV-1 and HCV are two of the most important blood-borne infectious agents. Hence, reliable, precise, and sensitive detection of these viruses in infected patients and donated blood units is highly important. Noticing the limitations of serological assays in detection of these infectious agents, this study was to use fast and sensitive molecular assays like real-time PCR.
Materials and Methods: In this trial, a home-brewed SYBR green-based multiplex real time PCR, on the basis of melting curve analysis, was developed for the single or simultaneous detection of HCV and HIV-1 infections in plasma samples. Data were analyzed using SPSS software version 16.
Results: The results obtained from different reactions on several clinical samples showed that the analytical sensitivities of the developed assay for HIV-1 and HCV were 200 and 100 copies/ml, respectively. It was also shown that the primers designed for each virus had no interaction with each other and other interfering agents.
Conclusion: Noticing the good level of sensitivity and specificity, easy handling, relatively low cost, and rapid analysis of samples, this method can be a useful and rapid approach for simple and effective detection of HCV and HIV-1 in plasma samples.
Behzad Khansarinejad, Mahdieh Mondanizadeh, Mohammad Rafeie, Siamak Mirab Samiee,
Volume 17, Issue 4 (7-2014)
Abstract
Background: The Real-time PCR assay has been established as the standard method for Human Cytomegalovirus (HCMV) quantitation in immunocompromised patients. However, the question of which one of whole blood or plasma specimens is better for viral quantitation is still unresolved for many clinical laboratories. To answer this question, the current study compares HCMV DNA load in whole blood and plasma samples.
Materials and Methods: In this prospective study, the whole blood and plasma samples were obtained from 41 transplantated patients and the viral load was detected using a validated, in-house Real-time PCR assay.
Results: Of the total 193 examined specimens, 174 were negative and 19 samples, from 16 patients, were positive in at least one of whole blood or plasma samples. Based on the results of linear regression analysis, the cytomegalovirus viral load was correlated in whole blood and plasma samples (R2: 0.872). However, the regression equation shows that the HCMV load in whole blood samples is higher than load of this virus in plasma. The validity of the quantitative results was confirmed by repeating the tests and analyzing the results using the repeated measure analysis.
Conclusion: Based on the results of the present study, HCMV quantitation in whole blood samples has a higher analytical sensitivity than in plasma samples.
Mina Zolfaghari, Behzad Khansarinejad, Ali Ganji, Zeinab Hamzehloo, Hamid Abtahi,
Volume 19, Issue 11 (2-2017)
Abstract
Abstract
Background: Ureaplasma and M. genitalium species belong to a kind of bacteria that are sexually transmitted and are the possible cause of pelvic inflammatory disease and nongonococcal urethritis, and et al. The aim of this study was to determine the urea plasma and Mycoplasma genitalium species frequency in women with vaginal infection and various sexual partners who referred to women, s health promotion and treatment center in Arak.
Materials and Methods: Endocervical swab samples from 110 women with vaginal infections referred to women’s health promotion and treatment center in Arak, were prepared. Patients’ personal information and identities during reception process were registered. The samples were transferred to the laboratory in the transport environment and after DNA extraction, were evaluated according to Real-time PCR assay.
Results: Urea plasma and Mycoplasma genitalium bacteria existed in 96(87.27%) and 4(3.63%) of patients, respectively. Among them, 4 cases had both bacteria infections. The amount of isolation in young women between 30-39 years old was more than others.
Conclusion: The results show that the colonization of urea plasma species in adult women is 40-80% and in studied group is 87.27%. These results indicate that with due attention to the increasing number of sexual partners and the increase of sexual activity, the urea plasma colonization of women will increase. In view of the potential influence of mycoplasma species on side effects resulted from pregnancy infection of mothers and mortality, on-time diagnosis and treatment will be increasingly essential.
Fariba Bani Talebi Dehkordi, Somayeh Reiisi, Asghar Bayati, Parisa Mohammadi Nejad,
Volume 20, Issue 3 (6-2017)
Abstract
Abstract
Background: Multiple sclerosis (MS) is a chronic inflammatory disorder described by central nervous system (CNS) demyelination and axonal damage. While the cause of MS is still unknown, it is extensively accepted that novel drug targets need to attention. Retromers are protein complex that have an essential role in endosomal trafficking, and retromer dysfunction has been associated to several neurological disorders. Therefore, this study aimed to compare the expression of SNX2 gene as a part of retromer complex in MS patients with health individuals.
Materials and Methods: In this case-control study, 50 samples of cases of multiple sclerosis (MS) and 50 healthy controls were enrolled. Followed verifying disease, 3cc peripheral blood was given from all subjects. Total RNA was extracted and complementary DNA (cDNA) was synthesized. The relative gene expression was determined using quantitative real-time RT PCR (qRT-PCR) and evaluated by 
method.
Results: The expression of SNX2 gene was lower in MS patients compared with healthy controls and it was statistically significant (p< 0.05).
Conclusion: Our study showed that the expression of SNX2 is lower in multiple sclerosis disorder. Considering the functional role of SNX2 as a protein involved in trafficking process, SNX2 may affect receptor function or drug targeting. Therefore, supplementary studies should be done to elucidate the exact mechanism of action of the gene in cellular trafficking.
Aref Mohammadipour, Najmeh Ranji, Leila Asadpour,
Volume 20, Issue 5 (8-2017)
Abstract
Abstract
Background: Pseudomonas aeruginosa is an opportunistic nosocomial pathogen that using several classes of antibiotics to treat has been led to the emergence of multiple drug resistance. One of the drug resistance mechanisms in Pseudomonas aeruginosa is overexpression of mexXY-oprM efflux pump system. Silybin as main flavonolignan of silymarin extracted from Silybum marianum is a hepatoprotective agent that its anti-bacterial properties was studied, recently. In this study, the effect of combination of silybin and ciprofloxacin on oprM gene expression in clinical isolates of Pseudomonas aeruginosa was evaluated.
Materials and Methods: In this study, seven ciprofloxacin resistant isolates of Pseudomonas aeruginosa were treated by ciprofloxacin (1/2MIC) only (control sample) and in the combination with silybin-encapsulated micelle (nanoparticles) (test sample). After 24h, RNA extraction and cDNA synthesis were performed in silybin treated and un-treated cells and oprM gene expression was quantitatively investigated by realtime PCR method.
Results: Results of this study showed that a silybin encapsulated in nanoparticles (400µg/ml) induces death up to 50% in resistant isolates treated by ciprofloxacin (1/2MIC) during 24h. Also, quantitative Real-Time PCR analysis revealed that silybin encapsulated in nanoparticles decreases the expression of oprM gene compared to silybin untreated cells.
Conclusion: It seems that Decrease of oprM expression in resistant isolates lead to decrease of mexAB-oprM and mexXY-oprM in cell surface, subsequently decrease of antibiotic withdrawal to extracellular environment and increase of sensitivity to antibiotics.
Marzieh Khoshbin Nazdik, Zeinab Khazaei Koohpar, Arezo Sayad,
Volume 20, Issue 6 (9-2017)
Abstract
Abstract
Background: Matrix metalloproteinases (MMPs) comprise a family of proteolytic enzymes.MMPs are capable of disrupting the blood-brain barrier (BBB), mediating the destruction of extracellular matrix and myelin components. Tissue inhibitors of metalloproteinases (TIMPs) are proteins which block the activitiy of MMPs. Matrix metalloproteinase-9 (MMP-9) facilitates T-cell migration into the CNS while the tissue inhibitor matrix metalloproteinase-1 (TIMP-1) inhibits MMP-9 actions. The aim of this study was to evaluate the expression of TIMP-1 gene (in RNA level) in blood cells of relapsing-remitting multiple sclerosis (RRMS) patients treated with IFNb.
Materials and Methods: In this study, the expression level of TIMP-1 gene was investigated in blood cells of MS patients compared to healthy subjects by Real-Time PCR.
Results: The RRMS patients manifested a lower expression level of TIMP-1 RNA than their normal counterparts, although the result was not significant (p=0.1).
Conclusion: The results of this study showed that there was no linear correlation between TIMP-1 expression level and risk of Expanded Disability Status Scale of Kurtzke (EDSS); nor was there any significant correlation between expression status of TIMP-1 and duration of the disease. Further studies are recommended to compare TIMP-1 RNA in patients before and after taking IFN-beta.
