Showing 44 results for 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.
S Rivandi , A Bahremand , S Zinali ,
Volume 56, Issue 6 (7-1998)
Abstract
In the present research work, a specific 285 bp DNA fragment was used for detection of Mycobacterium Tuberculosis complex. 100 samples were chosen randomly from sputum specimens that were negative with conventional methods (direct smear, culture, and radiometry), and examined by PCR 7 cases of them were positive. Also, 20 sputum specimens were obtained from suspected patients to tuberculosis, and examined by three methods (culture, radiometry and PCR). The sensitivity of PCR compared with culture and radiometry was 100%, the specificity of PCR compared with culture was 91.66%, and compared with radiometry was 68.75%. Therefore, results of PCR revealed, this method is more sensitive, specific and rapid and it can detect ycobacterial infectious agents within one day period.
Mirsalehian A, Jabalameli F, Kazemi B, Alizadeh S A,
Volume 61, Issue 6 (9-2003)
Abstract
Background: Staphylococci as a micro-organism, has the most importance to cause nosocomial infections, particularly in patients with indwelling catheters or other medical devices. Unfortunately 90% of Staphylococci isolated from the nosocomial infections are resistant to methicillin, and methicillin resistance strains are also resistant to a wide range of antimicrobial drugs, therefore detecting of these strains are valuable to eradicate the infection elements. Despite guidelines published by the national committee for clinical laboratory standards (NCCLS) for testing of susceptibility to methicillin for Staphylococci, the phenotypic method for detecting methicillin resistance remains controversial. Therefore, the genetic assays have been used to detect antimicrobial susceptibility of Staphylococci to methicillin.
Materials and Methods: Resistance to methicillin is coded by mec A gene in staphylococcus, and this gen must be detected in genetic assays. In this study 155 clinical staphylococcal isolates (70 coagulase- negative staphylococcus and 85 coagulase- Positive staphylococci) were evaluated for susceptibility to methicillin by using disk diffusion method.
ResuIts&Conclusion: Methicillin resistance was shown in 62 coagulase- negative staphylococcus (72.9%) and 27 coagulas positive staphylococcus (38.6%) but 63 coagulase negative Staphylococci (74%) and 28 coagulase positive isolates with mec a gene associated resistance were detected by PCR method. The results of this test were compared to the results for mec A gene detection by PCR test as a gold standard. The sensitivity, specifity and accuracy of the disk diffusion test for coagulase-negative staphylococcus were 96.8%, 95.45% and 96.47% and for coagulase positive staphylococci were 98.43%, 95.45% and 98.32% respectively.
Sedighi I, Rahimi H, Kakhodaee A, A Siadati ,
Volume 63, Issue 3 (6-2005)
Abstract
Background: Bacterial meningitis is a fatal disease with high mortality and morbidity that needs emergency management. But due to nonspecific signs and symptoms it&aposs diagnosis in children is difficult. Recently procalcitonin has been used for diagnosis of serious bacterial infections like bacterial meningitis. We conducted a prospective study in children for evaluation of procalcitonin in differential diagnosis of acute bacterial and viral meningitis.
Materials and Methods: In a prospective process research, we measured CSF procalcitonin levels in 43 children older than two months referred to Markaz Tebbi hospital. According to the results of universal PCR the patients were divided into two groups: bacterial meningitis (n=11) and nonbacterial meningitis (n=32). To analysis the results, Mann-Whitney test was used.
Results: CSF procalcitonin level in bacterial meningitis was significantly higher than viral meningitis (1.72±0.9 ng/ml and 0.71±0.04ng/ml respectively,Pvalue= 0.00). A serum procalcitonin level >0.5 ng/ml had high sensitivity and specificity ( 90.1% and 97.1% respectively) in the diagnosis of bacterial meningitis.
Conclusion: CSF procalcitonin level seems to be a valuable marker in differentiating between bacterial and viral meningitis.
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Volume 63, Issue 12 (4-2005)
Abstract
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Volume 63, Issue 12 (4-2005)
Abstract
R Kiani-Shirazi, S Zainali, M Karimipoor, B Zarbakhsh , R Alibakhshi,
Volume 64, Issue 2 (4-2006)
Abstract
Background and Aim:-thalassemia is the most common inherited disorder of hemoglobin (Hb) synthesis in the world. Alpha thalassemia most frequently results from the loss of one (- ) or both (- -) of the duplicated genes () on chromosome 16. Carriers of deletional forms of -thalassemia (-/- /-, or --/) are clinically normal but have a mild hypochromic, microcytic anemia. Compound heterozygotes (--/- ) called Hb H disease. Fetuses who inherit no genes (--/--) (Hb Bart&aposs Hydrops fetalis syndrome) die either inutero or shortly after birth, More than 95% of recognized -thalassemia involves deletion of one or both globin genes on chromosome 16.
Materials and Methods: The assay was tested on 114 Iranian individuals with low MCV and MCH levels but normal HbA2 who had not responded to Iron treatment. patients was referred to the Department of Biotechnology, Pasteur Institute of Iran by Health Centers. Genomic DNA was isolated from white blood cells by salting out method. We have developed a reliable, single - tube multiplex polymerase chain reaction (PCR) assay for the 7 most frequent - thalassemia deletions (-- SEA , --THAI, --FIL , -α20.5 , --MED, -α4.2 , -α3.7).
Results: DNA fromd thalassemia carriers was tested for the presence of different types of globin gene deletion (s). The - 3.7 and - 4.2 single gene deletions, and the Mediterranean (-- MED and - 20.5) double gene deletions were found in some samples.
Conclusion: The - 3.7 deletion was found to be the most common cause of globin gene deletion in our samples. Multiplex PCR for α gene deletion analysis is simple, rapid and sensitive.
E. Keyhani, N. Kohannia, N. Izadimood, M. R. Keyhkhaee, H. Najmabadi,
Volume 64, Issue 3 (5-2006)
Abstract
Background: Cervical cancer is the second leading cause of cancer death among women. In this cancer, the effects of prevention, early diagnosis and treatment more than other cancers decrease the mortality rate. In 1970 human papilloma virus (HPV) was introduction as major etiologic factor of cervical cancer. Different studies throughout the world revealed strong correlation between HPV and cancerous & precancerous changes in epithelial cells. Since cell culture and serological methods can not recognize the virus and its subtypes, the importance of the molecular methods including polymerase chain reaction (PCR) in early and definite diagnosis of virus is obvious.
Methods: In this study, after patient selection using the related protocol and completion of the questionnaires, 100 samples from cancer lesions of cervix selected. Then DNA extraction from paraffin blocks performed using standard method. Multiplex PCR with two pairs of primer (one as internal control) performed and the PCR product run on 8% polyacrylamid gel.
Results: The results showed that 73% of the tissues were infected by HPV.
Conclusion: This finding confirm the previous results based of correlation between HPV,and cervical cancer.
Aligholi M, Emaneini M, Hashemi F. B , Shasavan Sh, Jebelameli F, Kazemi B,
Volume 64, Issue 9 (9-2006)
Abstract
Background: Staphylococcus aureus (SA (is an important cause of nosocomial and community-acquired infections. The emergence of antibiotic resistance, especially in methicillin-resistant SA (MRSA) strains, has caused difficulties in treatment of such infections. The determination of antibiotic resistance patterns, particularly domestic patterns of Iran, is essential for appropriate treatment of MRSA infections and proper infection control measures in our country.
Methods: The antibiotic resistance of 338 SA isolates from various clinical specimens was determined by disk agar diffusion (DAD), minimum inhibitory concentration (MIC) and polymerase chain reaction (PCR) methods.
Results: Using the DAD method, 47% (160/338) of the SA isolates were resistant to oxacillin, and only 6% (20/338) were resistant to vancomycin. By PCR, 48% (162/338) of the isolates had the mecA gene. The MIC of oxacillin in 93% of isolates was higher than 256µg/mL. The MRSA isolates, showed a high resistant to gentamicin (40.5%), erythromycin (40%), and ciprofloxacin (38%). However, only a few of the SA isolates showed a high resistance to vancomycin (5%) or erythromycin (3.5%).
Conclusion: The results of this study can provide guidance for physicians toward a more appropriate treatment of SA infections in Iran, thereby preventing the emergence of further antibiotic resistance among SA. Our results also revealed the need for further investigations using a higher number of specimens representing a wider variety of locations to determine the antibiotic resistance patterns in our state more precisely.
Pajand O, Ziyaeyan M, Mousavi A, Hojabri Z, Kazemi B, Bahador A, Hamidian M, Mousavi A, Hashemi F B,
Volume 64, Issue 11 (10-2006)
Abstract
Background: Human Cytomegalovirus (HCMV) infections are a significant challenge in patients with Hematopoietic Cell Transplant (HCT). Acute Graft vs. Host (GVHD) is recognized as a predisposing factor for increased incidence of HCMV reactivation. Availability of rapid and accurate tests for HCMV detection in HCT recipients is of foremost importance in developing countries, such as Iran.
Methods: A total of 201 peripheral blood leukocyte (PBL) and plasma specimens from 26 allogeneic HCT recipients were examined for HCMV DNA by polymerase chain reaction (PCR) assay. Densitometric analysis of 257bp PCR products from clinical samples and 101-106 "cloned plasmid" per µg DNA containing a HCMV specific fragment were analyzed using LabWorks software (v3.0.02). Optical density of amplicons was plotted, and calculated HCMV viral loads were compared with the patients' antigenemia results.
Results: HCMV viral loads ranged between <102 to 1.35×102 copies per µg DNA among 7 HCT patients. In addition, 14 episodes of positive antigenemia assay in 7 patients in which peak HCMV load were compared with GVHD grade II-IV patients. Significant correlation was also detected between HCMV DNA load in PBL and plasma samples, as well as HCMV DNA load in PBL samples and antigenemia results. Receiver–Operating Characteristic analysis determined that 2,200 HCMV copies in PBL samples as the threshold value for initiation of Ganciclovir therapy.
Conclusion: This report shows that rapid and sensitive assays, like quantitative PCR, are extremely valuable for detection of active HCMV infection, and life-threatening HCMV disease in HCT recipients during the post transplant period. Furthermore, high HCMV DNA load among GVHD grade II-IV patients confirms the high risk of HCMV reactivation among these HCT recipients. Tests such as quantitative PCR also helps physicians initiate timely preemptive therapy and for a shorter period, which may lead to better clinical outcome in HCMV-infected transplant patients.
Shemshad K, Oshaghi Ma, Yaghoobi-Ershadi Mr, Vatandoost H, Abaie Mr, Akbarzadeh K, Telmadarraiy Z, Rafi-Nejad J, Derakhshandeh Paykar P,
Volume 65, Issue 1 (3-2008)
Abstract
Background: Malaria is still one of the main health problems in south and southeast provinces of Iran and recently on average 10,000-30,000 malaria cases were reported annually. Mosquitoes of Anopheles superpictus are one of the main malaria vectors in Iran and have been reported from all areas of the country including central plateau and plains of Alborz and Zagrous Mountains chains, and with low numbers in shore plains of the Persian Gulf and Caspian Sea. There are variations in larval and adult morphological characters and also in vectorial capacity of this species in different areas of Iran.
Methods: This study has been conducted to investigate rate of mtDNA variation among various populations of this species in Iran. The sequence variation of an 1512 bp length of mitochondrial DNA (mtDNA) cytochrome oxidase subunits 1 and 2 (COI-COII) and an 708 bp sequences of COI gene were analyzed by PCR-RFLP and PCR-direct sequencing respectively.
Results: This study showed that there are considerable variations between and within populations. Rate of variation was 12.3 % between populations and this was 2-5% for within Baluchistan population. Totally 4 haplotypes were observed between populations where 3 occur in Baluchistan and one in other places.
Conclusion: This is the first report on existence of various haplotypes in An. superpictus in science, and presumably this species comprising siblings and is a species complex. Further studies need to confirm this result and to determine the relationship between mtDNA haplotypes and their role in malaria transmission in each locality.
Shemshad K, Oshaghi1 Ma, Yaghoobi-Ershadi Mr, Vatandoost1 H, Abaie Mr, Zarei Z, Faghih Naini F, Jedari M,
Volume 65, Issue 8 (11-2007)
Abstract
Background: Anopheles superpictus is one of the main malaria vectors in Iran. The mosquitoes of this species are found throughout the Iranian plateau up to 2000 meters above sea level in the Alborz Mountains, south of the Zagros Mountains, and in the plains near the Caspian Sea and Persian Gulf. It has been reported that different geographical populations of An. superpictus play different roles in malaria transmission. Based on the presence or absence of a black spot/band on the apical segment of the female maxillary palpi, two morphological forms have been reported in this species. This work has been conducted to study other morphological features as well as the genetic structure of these two forms of An. superpictus in Iran.
Methods: The different morphological characteristics of 35 different populations were observed and recorded. An 887 bp portion of the mitochondrial DNA (mtDNA) cytochrome oxidase subunit I (COI) was amplified and assayed by restriction fragment length polymorphism (RFLP) using 18 enzymes and PCR-direct sequencing techniques.
Results: Among the morphological characteristics studied, there are significant differences between the two forms with regard to the length of the palp light band (p<0.01), wing length (p<0.5), and the distance from the branching point of the II/IV veins to the tip of the wing (p<0.05). Results also revealed that these two forms are sympatric in most localities of Iran. RFLP analysis and sequences of about 710 bp of the gene showed that there was great variation between and/or within the populations, but these variations were not associated with the morphological forms.
Conclusion: This is the first comprehensive study on the morphological and molecular characteristics of An. superpictus in the literature. To determine the role of these morphological forms or genetic haplotypes in malaria transmission, further molecular, cytological, morphological, and epidemiological studies are necessary.
Hajiabdolbaghi M, Allishah H.a, Rasoolinejad M, Bahador A, Izadi M, Mobaien A.r,
Volume 65, Issue 11 (2-2008)
Abstract
Background: Tuberculosis is still one of the most important causes of mortality and morbidity in many countries and is the second only to human immunodeficiency virus as a cause of death worldwide resulting from a single infectious agent. In 1993, the World Health Organization declared tuberculosis a global public health emergency. Conven-tional methods for the diagnosis of Mycobacterium tuberculosis (MTB) infections are time consuming, as MTB culture requires 3-8 weeks for growth. To determine the sensitivity of polymerase chain reaction (PCR) in peripheral blood mononuclear cells (PBMC), we have evaluated Mycobacterium tuberculosis DNA in peripheral blood samples with PCR technique in adults with new cases of pulmonary and extra-pulmonary tuberculosis. Setting: Department of Infectious disease of Imam Khomeini Hospital, 2004- 2005, Tehran, Iran.
Methods: In this cross-sectional study, we evaluated MTB DNA extracted from 3ml citrated peripheral blood samples from 95 adults with new cases of pulmonary and extra-pulmonary tuberculosis. DNA extraction was performed using a commercial PCR kit with IS1081 primers. For prevention of cross contamination and reduction of false positives, all steps were performed under laminar hood.
Results: The 95 patients, 59 of whom were male, had a mean age 44.44 years (SD±20.26) 69 cases had pulmonary and 26 had extra-pulmonary tuberculosis. PCR was positive in 32 (33.7%) patients and negative in 63 (66.3%) cases. The overall sensitivity and accuracy of the PCR assay was 44.1% for pulmonary, 19.2% for extra-pulmonary and 10% for disseminated tuberculosis, respectively.
Conclusion: The low sensitivity of the IS1081 primer MTB-PCR assay on PBMC may pose problems for the rapid diagnosis of tuberculosis. However, further studies are needed to confirm this technique as an alternative test for the diagnosis of tuberculosis.
Khalili M, Atapour M, Aali S, Azizollahi Ga, Azizollahi S,
Volume 66, Issue 2 (5-2008)
Abstract
Background: Chlamydia trachomatis (CT) is an obligate intracellular bacterium that causes genital disease and the most common sexually transmitted infection in the world. The most frequent risk factors associated with chlamydial infection are related to sexual behavior, multiple partners, and inconsistent condom use. Presenting primarily as urtheritis in men and cervicitis in women, CT a major cause of chronic pelvic inflammatory disease and subsequent infertility in women, eye and lung infection in newborns and other manifestations. Identification of CT-infected patients may prevent its spread and thereby reduce the high morbidity associated with CT infections. Polymerase chain reaction (PCR) is a sensitive and specific method for the detection of small quantity of bacterial DNA in clinical samples. The aim of this study was to determine the frequency of C. trachomatis by PCR in genital samples from patients in the city of Kerman.
Methods: A total of 130 genital samples including 64 endocervical and 66 urethral swab samples were collected by physicians. Nucleic acid was extracted from each sample using a commercial DNA extraction kit. PCR primers specific for a conserved region of the C. trachomatis omp2 gene, encoding an outer membrane protein, were used for amplification.
Results: A total of 9.2% (6.25% of cervicitis and 12.1% of urethritis) of the samples were found positive for CT using this PCR method.
Conclusions: The present study shows a high prevalence of CT infection, especially in men with urethritis. Such patients should be referred to genitourinary clinics for treatment and partner notification. Given its worldwide prevalence, further CT studies on more populations are needed to assess potential public health implications of these infections.
Mirsalehian, Feizabadi, Akbari Nakhjavani, Jabal Ameli, ,
Volume 66, Issue 5 (8-2008)
Abstract
Background: The resistance of Pseudomonas aeruginosa strains to broad spectrum cephalosporins may be mediated by extended spectrum b-lactamases (ESBLs). These enzymes are encoded by different genes located either on chromosome or plasmids. In this study, we determined the antimicrobial resistance patterns of P. aeruginosa isolates and screened for ESBL production.
Methods: After isolation from burn patients in Tehran Hospital, identification of P. aeruginosa isolates were assessed using biochemical tests. We then performed disk agar diffusion (DAD) according to CLSI guidelines to determine the pattern of antimicrobial resistance. The frequency of ESBLs and prevalence of the OXA-10 and PER-1 genes were determined with combined disk and polymerase chain reaction (PCR) methods, respectively.
Results: One hundred strains of P. aeruginosa were isolated. The resistance of these strains to cephpodoxime, aztreonam, ciprofloxacin, ofloxacin, ceftazidime, cefepime, imipenem, meropenem, cefotaxime, levofloxacin, piperacilin- tazobactam and ceftriaxon was 100%, 90%, 83%, 92%, 85%, 88%, 63%, 66%, 98%, 89%, 70% and 91%, respectively. Of these, 40 strains (40%) were ESBL positive, 29 strains (29%) were OXA-10 positive and 18 strains (18%) were PER-1 positive.
Conclusion: Our results confirm the need for proper antimicrobial therapy in burn hospitals, considering the resistance pattern and frequency of strains producing ESBLs and the presence of the OXA-10 and PER-1 genes. Since an increase in the prevalence of ESBL in P. aeruginosa strains might lead to the transfer of these ESBL genes to other gram-negative bacteria, we recommend the use of appropriate drugs, especially cephalosporins, in burn hospitals.
Chamani-Tabriz L, Tehrani Mj, Zeraati H, Asgari S, Tarahomi M, Moini M, Ghasemi J,
Volume 66, Issue 7 (10-2008)
Abstract
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Background: Chlamydia
trachomatis is a common and curable STI that may be
symptomatic or asymptomatic. The few studies on C. trachomatis among Iranian women have
had, for the most part, small sample sizes and are therefore unsuitable for
epidemiological deductions. The aim of this study was to estimate the
prevalence of urogenital C. trachomatis infections by PCR on urine samples of married women in their
fertile years in order to determine the need for a C. trachomatis screening program for
asymptomatic women in Iran.
Methods: This descriptive-analytical and cross-sectional study
was performed on 991 married women. The research material consisted of
questionnaires and urine samples, which were transported daily to Avesina
Research Institute, Tehran,
Iran, to
extract their DNA and prepare them for PCR tests. The gathered data were
analyzed by SPSS, version 13, and evaluated statistically by t-test, chi-square
test, Fisher's exact test and logistic regression, considering p<0.05 as
significant.
Results: Of all the subjects, 127 (12.8%) were positive by PCR for C. trachomatis.
The mean age of the participants was 28.88±
6.19 years. Infection was more prevalent among
those with lower levels of education, who were employed and not pregnant. This
infection was more prevalent among those who were using contraception, especially
condoms. Reproductive history revealed that infection was more prevalent among
participants with a history of vaginal discharge, pelvic pain, infertility and
low birth-weight infants, and less prevalent among those with a history of abortion,
preterm delivery and ectopic pregnancy. However, these patterns were not statistically
significant.
Conclusion: In populations with C. trachomatis prevalences higher than 4%,
screening programs are recommended. Thus, Chlamydia screening should be part of the health care program in Iran to reduce the
burden of this disease.
Mirhendi Sh, Adin H, Shidfar Mr, Kordbacheh P, Hashemi Sj, Moazeni M, Hosseinpur L, Rezaie Matehkolaie A,
Volume 66, Issue 9 (12-2008)
Abstract
Background: The clinical importance of yeast infections has increased in recent decades. There are 10-15 pathogenic Candida species. The current morphological and physiological methods for identification of Candida species are generally not easy to interpret and may be expensive or time-consuming. In the present study, we introduce and use a new approach for the identification and differentiation of medically important yeast species of Candida. In this method, size polymorphism of the internal transcribed spacer regions, ITS1 and ITS2, of the ribosomal DNA in various Candida species is used as the basis of species recognition.
Methods: The genomic DNA of 31 standard strains and 60 clinical isolates was extracted and PCR-amplified using two primer pairs (ITS1-ITS2 and ITS3-ITS4) separately. Both PCR products were mixed and analyzed after standard agarose gel electrophoresis. The species of the tested yeasts were identified by the electrophoretic patterns of the mixed PCR products of each sample, comparing the data obtained from the sequence analyses of ITS1 and ITS2 molecules.
Results: By this method, with the exception of C. albicans and C. dubliniensis, we were able to clearly differentiate nearly all common pathogenic Candida species, including C. albicans, C. glabrata, C. gulliermondii, C. parapsilosis, C. tropicalis, C. krusei, C. kefyr, C. lusinaniae and C. rugosa. All standard and clinical strains were identified correctly, without expensive methods such as sequencing and capillary electrophoresis.
Conclusion: It seems that the PCR-FSP method introduced in this study is the easiest molecular approach for the identification of a wide range of pathogenic Candida species and is applicable for diagnostic and epidemiological purposes in reference laboratories.
Noorbakhsh S, Tabatabaei A, Parvaresh M, Tonekaboni H,
Volume 66, Issue 10 (1-2009)
Abstract
Background: In recent years, many ill cases with cytomegalovirus reactivation in non-immuno compromised persons reported. Goal of study: to determine the CMV infection in cerebrospinal fluid of aseptic meningoencephalitis children hospitalized in Rasul & Mofid hospital (2005-2007).
Methods: In a cross sectional study 132 cases selected with simple sampling. CMV-DNA in their Cerebro spinal fluids searched by qualitative PCR.
Results: The age range of the study patients was 5 month- 13 years, median age= 2±3.7 years 87(65.9%) male and 45(34.1%) was female. The presenting signs and symptoms were convulsion 77(69.4%) meningitis 25(18.8%), loss of consciousness 47(37%) neurologic defects 15.9%. DNA extrated in 11 cases. Mycoplasma- DNA in 2cases DNA-CMV detected 2(1.5%). Positive DNA HSV found in 7(15.3%) of patients. DNA- HSV type- 15.3% (7/132) cases. An infant 5 month age with developmental delay, microcephaly and recurrent convulsions. A 1 year girl with brain atrophy and progressive hydrocephaly with intracranial shunt
Conclusions: Differentiation between herpes meningoencephalitis and other encephalopathy based on clinical signs in children is too difficult. CMV (1.5%) has lower rate than herpes simplex type-1 (5.7%). In addition to CMV and HSV1 all of herpes family viruses (varicella, herpes 6, 7, Epstein barr virus) could have role in children with meningoencephalitis. In recent years a sensitive, rapid, simple diagnostic test "Single tube Multiplex PCR" in cerebro spinal fluid recommend. Rapid diagnosis and faster treatment is necessary for decreasing mortality and morbidity in all of herpes meningoencephalitis cases
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.
