Measles, a viral exanthema of childhood, is the most contagious infection of humans. Measles virus is a member of the genus Morbillivirus of the family Paramyxoviridae. Despite the million"s lives saved by the global measles vaccination program of the World Health Organization (WHO) some 1 million infants and children continue to die each year from measles. In order to reduce the above mentioned figure more than 90% of children having 9 months of age or more must be immunized against measles.It is of great importance to do continuous serum surveys to measure herd immunity to disease.Several serologic tests are used to show the level of measles antibody, and each test has its own advantages and disadvantages.In this study it was attempted to apply indirect immunoFluorescent test (IIF), serum neutralization test (SNT), and plaque reduction test (PRT) to detect and measure measles antibodies. One hundred and twenty sera collected from children under the age of 5 living in some villages of Mianeh were tested with the IIF and SNT. Fifty of the sera were also tested using the PRT.To run the tests most of the reagent were used; e.g. fluoresceine-conjugated antihuman rabbit immunoglobulin measles antigen, and purified human IgG were first prepared.Taking the PRT as a gold standard test, the results obtainend from the IIF and SNT were compared with those of the PRT. Sensitivity and specificity of the IIF were equal to 84% and 67% respectively, whereas they were equal to 97% and 83% for the SNT.

Background and purpose: Rheumatiod arthritis (RA) is one of systemic autoimmune disease. In this disease, autoimmune antibodies are discovered, which have diagnostic and prognostic properties. One of them is APF, that can react with keratohyalin granules of buccal mucosal cell. Therefore, at first APF-IFA kit was designed and then antibody titer (Ab) was evaluated in RA patients.Materials and methods: Designing of kit was as follow: 1- Identification of satisfactory donors. 2- Antigenic substrate preparation. 3- Improvement of method and quality control.The method is based on the binding of APF to Perinuclear Keratohyaline line granules of buccal mocusal cells and its detection with total Anti Human Globulin (AHG) conjugated to FITC. After calibration of AHG, APF assayed in sera dilution of 1/5 , 1/10 to 1/1280 in three RA patients (52 RA patients mean age 48.0 ± 15.8), 23 sera of patient control groups (Mean age 32.5 ± 16.4) and 30 sera of healthy control group (mean age 32.1 ± 16.9).Results: From total of 52 RA patients (71.2%) were APF positive, in 2(8.7%) cases and in healthy control group 1(3.3%) case were APF positive.The designed APF kit had 98% accuracy by inter and intra assay methods. Sensitivity and specificity of APF in serum dilution was 71.2% and 94.3% respectively.Conclusion: According to the results of this study, 1/5 serum dilution has best diagnostic value for RA for best sensitivity and specificity, that means, 1/5 as a cut off, or significant minimum dilution in diagnosis and approving of RA, and considering 98% accuracy of designed kit, APF-IFA assay has medium diagnosis and high approving property for RA patients. Also Comparison between different groups under investigation showed that there is significant relationship between concentration of APF and Severity of disease.

Dear Editor: Leptospirosis is known as an emerging zoonotic infectious disease caused by the genus Leptospira. Several studies have shown that the incidence of the disease is significantly higher in tropical and subtropical areas. Some occupations, including veterinarians, rice farmers, butchers, fishermen, sewer maintenance workers, slaughterhouse workers, and laboratory staff, appear to be at higher risk for the disease. The aim of the present study was to investigate the epidemiology of leptospirosis in the Mazandaran province in northern Iran during 2012 - 2013. In this effort, 127 serum specimens were obtained from suspected individuals in different areas of the Mazandaran province during a period of eight months between November 2012 and July 2013. All 127 individuals were suspected of having leptospirosis according to physician diagnosis and the World Health Organization guidelines for diagnosis, surveillance, and control of the disease, including myalgia, neck stiffness, fever, headache, icterus, or a history of the disease prior to sampling.

Background: The IFA test is one of the most usual methods for detecting anti- Toxoplasma antibodies, although it has not any unique standardization. It seems that the microscopic judgment of results is an important confounder in IFA test. Therefore, we conducted the present study to clarify the role of microscopic observer, and other confounders on the test.Methods: Eighty sera were collected from patients suspicious to toxoplasmosis for detection IgG anti-T. gondii by this test. Samples were examined against different series of antigens, IgG antihuman conjugates, and observers.Results: There were no significant differences between the two series of antigens and conjugates.For the observers groups the kappa coefficient of the test results in the experts group (0.97, 0.94-1.00) were significantly higher than the less experienced observers (0.77, 0.68-0.87).Conclusion: We recommend the IFA test to be performed only in reference laboratories and by laboratory technicians that have enough experience for this test. Otherwise, we suggest the substitution of this test with other tests like ELISA for the diagnosis and epidemiological studies.

Background and Objectives: Nocardiosis is an acute or suppurative chronic disease caused by an aerobic, gram-positive, weakly acid-fast and soil-borne filamentous and organism. Nocardia asteroides which is the dangerous and most frequently pathogen, infects humans through the respiratory tract. The bacterium is primarily an opportunistic pathogen that causes the infection in patients with underlying immunodeficiencies.The main purpose of the study was to detect antibody titre against Nocardia in all study groups, using indirect immunoFluorescent assay (IFA). Correlation between the antibody titre against Nocardia with age, sex, occupation, and chronic pulmonary infection and corticosteroid therapy patients was also investigated.Material and Methods: The present investigation is a Cross-Sectional study conducted on a population consisted of 300 subjects including 200 hospitalized individuals' patients, nurses and healthcare workers from Imam Khomeini hospital, and 100 health adult blood donors. None of the patients had already been diagnosed to be affected by Nocardia.Results: Our results demonstrated four patients suffering from different infections, including TB, mycetoma, chronic pulmonary and chronic obstructive pulmonary diseases were IFA positive. None of the high risk hospital personnel, who were working in close proximity to the areas infected with Nocardia, were found to be IFA positive. Meanwhile there was no positive result in a group of patients (n=34) who were under corticosteroid therapy.Conclusion: Finally, considering the small sample size of the IFA positive cases no significant association between the IFA results and age, sex, occupation and clinical conditions of the subjects could be established.

Introduction: Wide and universal distribution of Legionnaires’ disease, abundance of reports about its prevalence and Pontiac fever from different countries, unresponsiveness to conventional antibiotic therapy in a number of pneumonia patients and the lack of any report about it in Iran directed us to perform this project.Legionnaires’ disease is seen in sporadic and epidemic form and the most prevalent cause is legionella pnuemophilia which produces a severe disease in vulnerable individuals with a high fatality rate (50-80%). It is also considered as one of the most fatal nasocomial infections.Methods: In the present research for isolating and detecting of L. pneumophila, bronchoalveolar specimens of patients were examined using culture and DFA methods. After 3-5 days of incubation at 37ºC, and humidity (about 95%), thin, convex and round colonies with blue-gray to blue-green appearance developed. Slides were prepared from these colonies and stained with Gram and Gimenez methods which displayed Gram – negative, small successive coccobacilli. Statistical analysis were performed by a software (SPSS, v10) using Fisher exact and McNemar Tests.Results: From among the 96 bronchoscopic specimens, 4 strains of Gram negative bacilli were isolated. Further investigations by means of biochemical tests and specific DFA revealed that they were L.pneumophila.In vitro drug resistance and sensitivity tests of Legionella showed that these bacteria were sensitive to Erythromycin, Riphampicin, Gentamycin, Doxycycline and Tubramicin, the sensitivity being more pronounced against Erythromycin than others. The above–mentioned organisms were resistant to Tetracycline and Ampicillin. The Fisher exact Test revealed that there is no correlation between frequency of L.pneumophila and the sex of patients (P value= 0.72). McNemar Test was performed and the results revealed that there is no significant difference between the results obtained by culture and DFA tests (P value= 1).Conclusion: L.pneumophila has no correlation with the sex of patients. According to the results obtained by McNemar test, DFA method can be used for rapid detection of L.pneumophila, although further studies with larger quantity of specimens need to be performed to substantiate results.

Introduction: This study aimed to determine diagnostic value of immunoperoxidase in comparison with immunofluorescence in the diagnostic assessment of kidney biopsy specimens.Materials and Methods: Forty-eight kidney biopsy specimens were used to compare a direct immunofluorescence technique with immunoperoxidase techniques on paraffin sections. The sensitivity and specificity were calculated. The kappa statistic for agreement between the two tests was categorized as poor (zero to 0.2), moderate (0.21 to 0.45), good (0.46 to 0.75), and almost perfect concordance (0.76 to 1.0).Results: Compared with immunofluorescence, the immunoperoxidase technique presented a sensitivity of 88.55% and a specificity of 69.22%. Its sensitivity in the staining for IgG, IgM, and IgA was 93.75%, 95.45%, and 76.47%, respectively. The specificity of this test in the staining for IgG, IgM, and IgA was 54.54%, 57.14%, and 96.00%, respectively. The overall kappa value was 0.60 and it was 0.60 for assessing staining intensity. There was a moderate agreement between immunoperoxidase and immunofluorescence in the positive or negative staining for IgG and IgM, as well as a good agreement in the positive or negative staining for IgA. For the staining intensity, the two tests had a good concordance for IgG and IgA and a moderate concordance for IgM.Conclusions: Although immunoperoxidase method has a lower overall diagnostic performance as compared to immunofluorescence, given the good concordance between the two techniques, it can be an alternative method for immunofluorescence study of kidney biopsy specimens, particularly where immunofluorescence fails or is not available.

Objective: This study was carried out in order to produce FITC conjugated antibody against pestivirus antigens to detect pestiviral infection. This study was begum on Jan. 2000 and it was ended on Oct. 2001.Design: Field experiment.Procedure: At first 10 rabbits were immunized with NADL strain of BVDV. The immunoglubulins were conjugated with FITC and were passed on sephadex G25. To obtain a desirable titer of conjugatedsera different dilution of antibody in presence of positive and negative samples a were used.Statistical analysis: Mcnemar and Chi square tests.Results: As a result, a titer of 1/20 of conjugate turned out to be desirable .At the last term of study from 210 selected cell culture tube, 140 of them were infected with BVD virus and 70 were kept as control. The results of FA microscopy were indicated a coordination of about %100 between standard (Biox) and our conjugated sera. On 81 fields infected samples examined by 2 antisera, only one sample was negative by our prepared conjugate, although this sample was positive by standard conjugate.

Background and Objectives: The inability of classic fluorescence-activated cell sorting to single cancer cell sorting is one of the most significant drawbacks of this method. The sorting of cancer cells in microdroplets significantly influences our ability to analyze cancer cell proteins. Material and Methods: We adapted a developed microfluidic device as a 3D in vitro model to sorted MCF-7 cancer cells on a chip. A prefabricated microfluidic droplet chip was used in this research. Then, with the help of synthesized Fluorescent probes, MCF-7 cancer cells were separated from normal cells. Results: This research presents a modification of GQD bead for high-throughput analysis and sorting single cancer cells. We elaborate a binding assay as an example of this approach for detecting MCF-7 cancer cell lines. Graphene quantum dot-decorated mesoporous silica nanoparticles (GQD@MSNPs) act as Fluorescent optical beads coated in microfluidic droplets. The Fluorescent beads capture cancer cells. To enable droplet sorting at 200 Hz and cell enrichment, a measurable fluorescence signal is generated when cancer cells bind to these beads and boost the drop's fluorescence emission. Conclusion: Herein, we report in vitro results showing that the as-prepared GQD@MSNs have exceptional luminous characteristics. The specific surface area and pore volume of GQD-MSNs were found to be 50% and 40% higher than those of pure MSNs, which is rather remarkable. Because of these improved qualities, GQD@MSNs are demonstrated a large sorting capacity that makes them ideal for diagnosis.

Objective: Confocal laser scanning microscopy (CLSM) is relatively a new light microscopical imaging technique with a wide range of applications in biological sciences. The primary value of CLSM for the biologist is its ability to provide optical sections from a three-dimensional specimen. The present study was designed to assess the thickness and content of in vivo accumulated dental plaque using CLSM and scanning electron microscopy (SEM).Materials and Methods: Acroflat lower arch splints (acrylic appliance) were worn by five participants for three days without any disturbance. The formed plaques were assessed using CLSM combined with vital fluorescence technique and SEM.Results: In this study accumulated dental plaque revealed varied plaque microflora vitality and thickness according to participant’s oral hygiene. The thickness of plaque smears ranged from 40.32 to 140.72 mm and 65.00 to 128.88 mm for live (vital) and dead accumulated microorganisms, respectively. Meanwhile, the thickness of plaque on the appliance ranged from 101 mm to 653 mm. CLSM revealed both dead and vital bacteria on the surface of the dental plaque. In addition, SEM revealed layers of various bacterial aggregations in all dental plaques.Conclusion: This study offers a potent non-invasive tool to evaluate and assess the dental plaque biofilm, which is a very important factor in the development of dental caries.