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Showing 13 results for Elisa

Fooladsaz K, Ansari M, Javad Rassaie M,
Volume 62, Issue 1 (4-2004)
Abstract

Background: Migraine is a chronic hereditary and relapsing headache. With regard to the prevalence of this ancient disease and its economic complications in country, in this study , nocturnal serum melatonin of migraine patients and control subjects have been evaluated and compared by ELISA kit.

Materials and Methods: Fifty migraine patients (mostly women) were compared to a control group (mostly men) matched according to age. Results: Statistical analysis revealed a decrease in nocturnal serum melatonin levels for migraine patients (32.9 28.4) compared to the control one (75.6 56.8). With using of t-test by ELISA kit showed significant difference (p=0.0064).

Conclusion: With regard to this, the pineal gland has the main role in the synchronization of the organism with the environmental conditions and migrainous headaches.


Salehi Nodeh A.r, Ghaffori Sh, Alimohamadian M.h, Sarraf Nejad A, Mirshafiei A,
Volume 64, Issue 11 (10-2006)
Abstract

Background: TPS is one of the tumor markers which has specially been considered due to its exclusive physiological characteristics like its easy measurement in serum of cancer patients. This study has been due to evaluate the efficiency of this tumor marker in the prognosis, treatment control and follow up of patients with gastrointestinal cancers including esophagus, stomach and colorectal.
Methods: TPS has been measured in 109 persons including 28 healthy people and 81 patients with different gastrointestinal malignancies which were composed of 38 patients with esophageal cancer, 20 ones with stomach cancer and 23 ones with colorectal cancer. Sampling has been done in three times depending on treatment methods. TPS has been measured with ELISA in samples which contend of 2 to 3 ml of serum from patients and the health.
Results: The obtained results, demonstrate the obvious changes in TPS serum level in patients underwent various treatment procedures.
 Conclusion: The results have revealed that the serum TPS is not only as a measure of prognosis but also would be helpful in follow up and treatment control of the disease. Moreover the results has shown that serological analysis can be settled in the diagnosis and follow up with production of polyclonal antibody against TPS gene family and planning appropriate pattern.
Moazeni M, Gaur S.n.s,
Volume 65, Issue 11 (2-2008)
Abstract

Background: Fasciolosis is a worldwide disease with major economic and public health consequences. Early detection of the infection is important for the prevention and control of the disease. ELISA allows for early detection of fasciolosis in man and animals. Fasciolosis is caused by Fasciola hepatica and F. gigantica in man and domestic animals respectively. These two species have many similar morphological characteristics. In this study, the crude antigens of these two species are investigated by ELISA test.

Methods: The excretory-secretory and somatic antigens of two species were prepared from adult flukes collected from the bile ducts of sheep and stored at -20oC. For the preparation of the antisera, the antigens were injected to laboratory-bred rabbits. Each rabbit received five injections at intervals of seven days, starting with 0.5 ml and ending with 2.5 ml. Ten days after the last injection, the rabbits were bled, and serum samples separated and stored at -20oC. The reaction between homologous and heterologous antigens and antisera was tested by ELISA and optical densities were recorded.

Results: Excretory- secretory and somatic antigens of each species showed a strong positive reaction with the antisera of the other species. In a homologous combination of antigens and antisera, a stronger reaction was observed compared to the heterologous combination, therefore many antigenic materials of both species are the same.

Conclusion: The differences of these crude antigenic materials of F. hepatica and F. gigantica are insufficient to prevent cross reaction of two species by ELISA. Further investigations are recommended for the identification, detection and purification of antigenic material of each species to improve the specificity of this assay.


Khalvat A, Rostamian A, Najafizadeh S R, Movasseghi S,
Volume 65, Issue 11 (2-2008)
Abstract

Background: Rheumatoid factor (RF) is an IgM antibody against the Fc portion of IgG, which together form an immune complex. RF is an important criterion in the diagnosis of early-stage rheumatoid arthritis (RA) and prognosis of RA pathogenesis, as higher levels of RF indicate a higher possibility of more damage. Although 2/3 to 3/4 of patients that undergo ordinary standard tests and have final clinical diagnosis are also positive for RF, a 70-90% prevalence of RF among RA patients can be achieved, depending on the method of detection and the target antibody, IgG or IgM. In this study, we measured the frequency of IgG and IgM RF isotypes using the ELISA and latex agglutination methods and compare these results with those of a hospital control group, tested using standard methods, in order to determine the best method for the measurement of RF.

Methods: Of the patients referred to the Rheumatology Clinic of Imam Khomeini Hospital during 2005-2006, one hundred randomly selected rheumatoid arthritis patients, 75 females and 25 males, with classical or definite rheumatoid arthritis (defined by the criteria of the American College of Rheumatology), with a short disease duration of 12-24 months, underwent testing for RF using the latex method for IgM and ELISA for IgM-IgG. The healthy control group (75 females and 25 males) were tested for RF using the ELISA method for IgM-IgG. The variables were compared using the Pearson's chi-square test.

Results: We found that the measurement of RF among RA patients using did not differ significantly between the two methods. The immune complex in RA is mainly IgM. The positive IgM results in RF patients using two similar methods showed a significant relationship by Pearson's correlation co-efficient (r=0.60, p<0.001). In addition, comparison of the IgM and IgG RF by ELISA showed a weak correlation with low significance (r=0.10, p<0.001). In sum, this study showed a significant difference (r=0.24, p<0.001) between the IgM in RA patients and that in healthy people, who had no IgM or IgG RF.

Conclusion: Approximately 75% of confirmed RA cases had the IgM RF however, we found little advantage in using the one method over the other, nor was the measurement of IgG more useful than IgM as a diagnostic criteria.


Gholampour Azizi I, Khoushnevis S H, Hashemi S J,
Volume 65, Issue 13 (3-2008)
Abstract

Background: Aflatoxins are severe toxic secondary metabolites found in most plant products. When animals consume contaminated feed stuff to Aflatoxin B1 (AFB1), the toxin is metabolized by liver and is excreted as Aflatoxin M1 (AFM1) via milk. Aflatoxins are acute toxic compounds, immunosuppressive, mutagen, tratogen and carcinogen.
Methods: During the winter of 2006, pasteurized and sterilized (ultra high temperature) (UHT) milk packages were collected from supermarkets in Babol city. 78 pasteurized and 33 sterilized milk, totally 111 samples were tested for AFM1 by competitive Enzyme Linked Immunosorbent Assay (ELISA). Solid phase in plastic micro wells coated whit anti-Aflatoxin M1 antibodies. We added 100 microliter skimmed milk and Aflatoxin M1 standard solutions in each well. In each plate, we appointed seven wells for standards. Plates were incubated at 20-25° centigrade for 45 min. Each well was washed four times by washing buffer 20X concentration. Then 100 micro liter conjugated solution (100X) was added to each well, and the plate was incubated at 20-25 centigrade for 15 min. After that, the wells were washed. After adding the substrates to wells, we incubated the plate at 20-25° centigrade in a dark place for 15 min. The reaction was stopped by stop solution. After one hour, light absorption was read at 450 nm by ELISA reader.
Results: AFM1 were detected in 100% of all samples. 100% of samples were above of European community regulations (50ng/l). AFM1 contamination mean levels pasteurized and sterilized milk were 230.5 and 221.66 respectively. Therefore more than four fold levels European community. There is not a significant relationship between AFM1 contamina-tion level and different months of winter applying statistical test.
Conclusion: The results showed the need for introducing safety limits for AFM1 levels in child milk under Food Legislative liable of Iran. Aflatoxin M1 contamination is a serious problem for public health, and it is potentially hazardous for human health.
Asadi Amoli F, Akbarzadeh Jahromi M, Attar M, Azadbakht F,
Volume 66, Issue 8 (11-2008)
Abstract

Background: Chlamydia Trachomatis is the most common cause of trachoma and subsequently give rise to neonatal chlamydial conjunctivitis (NCC), adult ophthalmic inclusion infection, sexually transmitted diseases (STD) and pneumonia. The goal of this study was to access the incidence of chlamydia trachomatis in the normal (ophthalmic infection free) population.

Methods: In a cross sectional study 250 patients referring to Farabi Eye university Hospital Tehran, Iran for non infectious ophthalmic disease in different age categories were selected and accessed for chlamydial IgM and IgG by ELISA method.

Results: 250 patients (50% men and 50% women) with the mean age of 40 (ranging from one to 83 years old) were tested. IgG was detected in 11 (five females and six males) patients (4.4%) All of them had more than 31 years old. IgM was detected in 18 (13 females and 5 males) patients (7.2%). No test revealed simultaneous high IgG and IgM titre in the same patient.

Conclusions: There was a low grade of chlamydial infection in our study population. So it is recommended to use serological methods for screening of ophthalmic infections in centers where no other test methods are available and in case of positive results confirmatory antigen tests to be used.


Mohammadi M, Mirjalili A, Habibi Gh, Falahi Sh, Sarafnejad A, Eatemadi A, Boutorabi Sm,
Volume 67, Issue 1 (4-2009)
Abstract

Background: Anti-dsDNA antibodies frequently found in the sera Systemic Lupus Erythematosus patients, particularly in active disease stage. Nowadays exploit different eukaryotic and prokaryotic dsDNA as antigen source and different reagents as binder. The aim of this study to compared two dsDNA different sources and tow different kinds of reagents for binder in ELISA test.

Methods: In this study bacterial genomic DNA from E.coli (ATCC 25922) and genomic DNA from calf thymus extracted with high purity and were used as antigens for IgG anti-dsDNA detection by ELISA. To coat dsDNA in microtiter wells, tow different kinds of reagents including methylated -BSA and poly-l-lysine (for pre-coating) are used. Sera from systemic lupus erythematosus patients and from normal blood donors are used to assess sensitivity and specificity of our ELISA test in compared with IF test and commercial kits.

Results: Our results displayed pre-coating of microtiter plates with methylated -BSA reduce nonspecific binding reaction and the relative sensitivity and specificity of ELISA increased when calf thymus DNA is employed as antigenic source in compared with IF test and commercial kits 80%, 88% and 100%, 98% respectively, but when E.coli DNA is used 73%, 69% and 85%, 79%, respectively.

Conclusion: The genomic DNA from calf thymus is a potentially useful source of antigen for detection of anti-dsDNA by ELISA. Also the use of methylatted- BSA could have an effective role in reducing of nonspecific binding reactions.


Abdolreza Soudbakhsh , Habibollah Mortazavi , Mahbobeh Hajiabdolbaghi , Mehrdad Hasibi , Sirous Jafari , Hamid Emadi Kochak, Esmaili Djavid,
Volume 67, Issue 6 (9-2009)
Abstract

Background: Finding a reliable diagnostic method for brucellosis is the most challengeable problem. In this study we determined the optimal diagnostic cut-off point for ELISA test.
Methods: We gathered 56 confirmed cases of brucellosis. Furthermore blood samples from 126 controls including 73 healthy controls and 53 without brucellosis febrile patients were collected. In all of the cases and controls ELISA Ig G and ELISA Ig M levels were measured and compared with each other by Box plot graph and the Receiver Operating Characteristic (ROC) curve. The sensitivity and specificity of ELISA Ig G and Ig M were fixed in different cut-off values and Ig G and Ig M levels yielding maximal sensitivity plus specificity were selected for determination of optimal cut-off point.
Results: The nineteen patients had positive blood cultures for Brucella melitensis. The standard agglutination test results were 1/160 or more in 54 patients. The Box plot graph indicated a high degree of dispersion for Ig G and Ig M data in patients with brucellosis compared with febrile patients without brucellosis and healthy controls. We observed partial overlap for Ig M data (not for Ig G) between cases and controls. The area under ROC curve for discrimination of cases and healthy controls was more for Ig G than Ig M.
Conclusions: The ELISA Ig G is more reliable test than ELISA Ig M in diagnosis of brucellosis. Using cut-off of 10 IU/ml and 50 IU/ml have the most sensitivity (92.9%) and specificity (100%) for ELISA Ig G test, respectively.

Pourakbari B, Mamishi S, Pajand O, Nadji Sar, Mahjob F, Kochakzadeh L, Izadyar M, Parvaneh N, Saboni F,
Volume 67, Issue 11 (2-2010)
Abstract

Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 Background: Latent Epstein- Barr virus (EBV) genomes are found in the malignant cells of approximately one-third of Hodgkin's lymphoma (HL) cases. Detection of EBV viral DNA could potentially be used as a biomarker of disease activity. Our goal was to compare of EBV DNA detection in samples obtained from lymphoma patients versus controls.
Methods: One milliliter uncoagulated and 1ml coagulated blood sample for DNA extraction and serum analysis using ELISA for IgG anti EBNA-1 were obtained from 44 lymphoma patients and from 44 normal controls, respectively. EBV genome, EBNA-2, was examined from DNA extracts of paraffin embedded and blood samples using Nested PCR with type specific inner primers.
Results: Positive results for ELISA, Blood and biopsy PCR in study group were, 84.1%, 27.3% and 13.6%, respectively. However, these results in control group were 47.7% and 16% for ELISA and Blood PCR assays, respectively. Positive results in ELISA, Blood PCR and Biopsy PCR in Hodgkin and non-Hodgkin patients were found in 21(84%), 6(24%), 4(16%) and 16(84.2%), 6(31.6%), 2(10.5%) of specimens, respectively. No significant differences in EBV detection were found between these two patient groups (p values for ELISA, Blood PCR and Biopsy PCR were 0.26, 0.73 and 0.68, respectively).
Conclusion: Comparison of ELISA and Blood PCR results in children and adult patients with the same age of controls have showed difference in ELISA results of children, only. None of the test results have showed statistically significant difference between Hodgkin and non-Hodgkin patients. However, the mean of ELISA results in Hodgkin patients was higher as compared with controls. Blood PCR assay cannot be recommended as a biomarker of disease activity in EBV positive Hodgkin's lymphoma patients.


Kardar Gha, Pourpak Z,
Volume 68, Issue 3 (6-2010)
Abstract

Background: The hypersensitivity to cow’s milk allergens is the most common allergies in children at the first year of life. The specific IgE evaluation is one of the important methods in diagnosis of allergic disease. The aim of this study was development of a sensitive and credible procedure for detection of cow’s milk allergens specific IgE. Methods: The allergen discs were prepared by coating of allergens on nitrocellulose paper. After incubation of allergen discs with patients serum, anti-human IgE conjugated were used. In following optimization of any step of ELISAs test, a complete kit was designed. Efficiency of designed kits were evaluated by determination of specific IgE in normal (n= 29) and patient (n= 153) children serum samples and compared with commercial kits. Results: The specific IgE against three allergrns involving casein, -lactalbumin and - lactoglobulin were measured on normal and patient children serum with designed and commercial ELISA kits. Results were demonstrated specificity of 93%, 89.7% & 82.8% and sensitivity of 86.3%, 81.3% & 89.6% respectively for casein, -lactalbumin and - lactoglobulin specific kits and these results were similar and comparable with commercial kits. Conclusion: The Designed kits in comparison with the commercial kits were showed equivalent sensitivity and specificity. The designed kit stability was ultimately one month, probably due to don’t using of stabilizers for prepared allergen discs. We suggest these kits for commercial product in Iran and we hope be helpful for easier accesses for Cow’s milk allergy diagnosis and extend that for other allergens.
Noorbakhsh S, Shekarabi M, Kalbasi Z, Tabatabaei A, Tonekaboni H, Afsharkhas L, Vafaei-Shahi M,
Volume 68, Issue 5 (8-2010)
Abstract

Background: M. pneumoniae infection in children is usual and diagnosis of its neurologic complications for rapid treatment is very important. To compare the CSF- M. pneumoniae antibody level between febrile children with acute neurologic signs (Menigoencephalitis, Guillan Barre Syndrome (GBS), Transverse myelitis, Ataxia and so on) with unaffected ones.

Methods: A cross sectional/ case control study in pediatric wards of Rasoul-e-Akram & Mofid hospitals (2007-2009) was done. The amount of Specific M. pneumoniae IgG (ELISA) antibody level determined in CSF of 55 cases and in 10 controls. Chi square values (CI 95%, p< 0.05) calculated for all categorical variables. Sensitivity specificity Positive Predictive Value (PPV) Negative Predictive Value (NPV) of CSF antibody level determined by using the Area under the ROC Curve.

Results: Cases (n= 55) aged between five month to 13 years with mean age of 3.84±3.43 years. Area Under Curve (AUC) in ROC was 0.876 (%95 CI, 0.78- 0.96 p< 0.0001). Cut off level for antibody was 0.0025 with 73% sensitivity 90% specificity 100% PPV 28.8% NPV. CSF antibody level had significant difference between cases and controls [0.08± 0.26 Versus 0.001± 0.001 p: 0.02] It had poor agreement between cases and controls (Kappa= 0.27). Lowest amount seen in cases with aseptic meningitis highest amount observed in cases with GBS and cases with focal neurologic signs.

Conclusion: The presence of very low amount (0.0025) of M. pneumoniae antibody in CSF of febrile children with acute neurologic signs had 70% sensitivity and 90% specificity 100% PPV but had low (28.8%) NPV. M. pneumoniae would be a rare cause in cases with aseptic meningitis. Finding the M. pneumoniae-DNAs in CSF are not so frequent (2%) but in high suspicious cases adding this test to determining the CSF antibody level might be helpful.
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.


Moradkhani S, Mahdi Mm, Daneshvar H, Bazargan Harandi N, Baneshi Mr,
Volume 70, Issue 10 (1-2013)
Abstract

Background: Atopic dermatitis (AD) is one of the most common chronic, highly pruritic and inflammatory skin diseases. The exclusive influence of breastfeeding in the prevention of inflammatory diseases is a matter of debate. In this study, we aimed to determine the concentration of interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), interleukin-13 (IL-13) and interleukin-4 (IL-4) cytokines as anti Th2 or anti Th1 cytokines in breast milk and their relationship with atopic dermatitis in breastfed infants.
Methods: This study carried out in Afzalipour Hospital of kerman during one year from 2010 to 2011, we selected 50 breastfed infants with AD as cases and 50 healthy infants without AD or any other allergic disease as the controls. The concentrations of pro- and anti-inflammatory cytokines were measured by ELISA in the mothers' milk. The demographic characteristics were recorded in a data collection form. Moreover, severity of the disease was determined by SCORAD index. T-test and logistic regression were used for assessment of the correlation among study variables.
Results: The concentrations of IFN-γ and IL-13 were significantly higher (respectively, P=0.04, and P=0.02) in the case group. However, logistic regression revealed that only IFN-γ significantly increased the risk for atopic dermatitis (P=0.02). Concentration of TNF-α was similar in the milk from mothers belonging to the two groups.
Conclusion: The results indicate that the concentrations of IFN-γ, IL-13 and IL-4 cytokines are higher in the milk of mothers whose infants have AD. However, the risk for atopic dermatitis increases by 49% by every ten-unit (in pg/mL) increase in the level of IFN-γ.



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