Diploid and continuous cell lines are used to propagate viral vaccines. At Human Viral Vaccine Department of Razi Vaccine and Serum Research Institute, MRC5 diploid cell is used for the development of live attenuated measles, mumps, rubella, and three types of poliovirus vaccines. Additionally, three continuous cell lines (i. e., RK13, HeLa, and Vero) are applied in quality control tests. Accordingly, cell cross-contamination can occur at cell culture labs, hence controlling the identity and specificity of cells is essential. INDIRECT IMMUNOFLUORESCENCE is a sensitive, specific, and simple test for cell identification. The present study was designed to develop the inhouse INDIRECT IMMUNOFLUORESCENCE test (IIF) as follows: homemade polyclonal anti-MRC5 serum was prepared in rabbits, and cross-reactive antibodies to RK13, HeLa, and Vero cells were eliminated. The diploid and continuous cell lines were fixed on Teflon slide using cold methanol and acetone. The reproducibility of the inhouse IIF test was evaluated using the agreement Kappa test. The purity of the three batches of MRC5 working seed cell at Human Viral Vaccine Department of Razi institute was verified using IIF and no contamination with continuous cell lines was detected.

Background: IMMUNOFLUORESCENCE and serology analysis are the most common laboratory methods for diagnosing antinuclear antibodies in autoimmune diseases and are paramount for screening and therapeutic purposes. This study aims to estimate the sensitivity and specificity of antinuclear antibodies measured by automated INDIRECT IMMUNOFLUORESCENCE and enzyme-linked immunoassay in patients at risk for autoimmune diseases. Methods: Serum antinuclear antibodies in 3020 patients suspected of autoimmune diseases at Nobel Medical Laboratory, Esfahan, IRAN, were measured from 2017 until 2020 with automated INDIRECT IMMUNOFLUORESCENCE and enzyme-linked immune assay methods. The sensitivity, specificity, prevalence, positive and negative predictive value, and likelihood ratio were calculated for each technique. In addition, the receiver operating characteristic curve (ROC) was analysed as a statistical method for assessing the diagnostic accuracy of these tests. Results: The IMMUNOFLUORESCENCE method demonstrated low sensitivity and high specificity compared with the enzyme-linked immunoassay. For the automated INDIRECT IMMUNOFLUORESCENCE method, sensitivity and specificity were 88% and 62%, respectively, whereas this number for the ELISA method was determined as 89.6% and 28.5 %, respectively. Conclusion: It is crucial to choose a suitable method for detecting autoantibodies for diagnostic purposes. ANA analysis by a sensitive test, such as an enzyme-linked immunoassay, should be used for screening. In contrast, a highly specific test, such as an INDIRECT IMMUNOFLUORESCENCE assay, should be used to confirm the result and monitor dynamic treatment.

Background: Vulvovaginal candidiasis occurs due to the overgrowth of candida in genital system mucosa of females. Symptoms and signs of vulvovaginal candidiasis are unspecific, therefore diagnosis is not certain. The aim of this research was comparison of the result of INDIRECT IMMUNOFLUORESCENCE and ELISA with culture and direct microscopy examination in patients with vulvovaginal candidiasis.Methods: This was a comparative-descriptive study performed on 87 patients and 50 normal cases as controls. All specimens were examined using direct microscopy, culturing and complimentary test to differentiate the candida species from each other. Serological tests such as INDIRECT IMMUNOFLUORESCENCE and ELISA were performed on sera of the patients. To compare the quantitative and qualitative data, t student test, chi-square and exact fisher test were used, if necessary.Results: Out of 87 specimens, 50 cases were diagnosed as vulvovaginal candidiasis. The most frequent isolated pathogens were C.albicans, C.glabrata, C.kefyr, C.inconspicua and C.famata, respectively. Also, in control group, the most frequent pathogens were C.albicans, C.glabrata and C.kefyr, respectively. In this study all of normal cases were negative in INDIRECT IMMUNOFLUORESCENCE test and in patients group 42 person (84%) were positive and 8 (16%) negative. Control participants were negative in ELISA and in patient group 40 person (80%) were positive and 10 person (20%) negative.Conclusion: It seems in cases that direct microscopic and culturing methods is impossible, ELISA and INDIRECT IMMUNOFLUORESCENCE can be used as an alternative method.

Leptospirosis is a worldwide zoonotic disease with a much greater incidence in tropical and temperate regions. It has now been identified as one of the emerging infectious diseases. Since, the incidence of Leptospirosis is significantly high in temperate regions among rice field workers and livestock animals at Mazandaran province, it is neccessery to evaluate condition of Leptospirosis that will help to recognize the most prevalent for vaccine development. In this survey, 127 serum samples of suspected human were collected from different parts of Mazandaran during eight months (during november 2006 to july 2007) and were tested by INDIRECT immunoflurescence assay (IFA) in Amol Branch, Institute Pasteur of Iran. To recognize common pathogenic serotypes, collected samples were tested using Microscopic agglutination test (MAT) in Leptospira Research Laboratory, Faculty of Veterinary Medicine, University of Tehran. Prevalence rate of Leptospirosis in Mazandaran province was 58.26% using IFA and MAT. In this survey, 74 samples were positive which 82.43% of them were belonged to male and 17.52% to female. Rice field workers showed the most frequent occurrence of infection, so among positive cases 66.22% were to the rice field workers and showed significant difference with other occupations (P=0.007). Among 107 samples that were collected from males and 20 from females, 57% and 65% samples were positive, respectively. There was no significant difference between gender and Leptospirosis (P>0.05). Living place (urban or rural) and contact with animals did show no significant difference with Leptospirosis. The most positive cases were found in age range of 40-50 years (25.67%). The most prevalent serotypes which were recognized in Mazandaran province using MAT, included, Ballum (16.77%), Sejroe (14.29%), Tarassovi (13.64%), Australis (11.69%), Pyrogenes (7.79%), Javanica (6.5%), Icterohaemorrhagiae (6.5%). Using a MAT titer of ³ 1:100 as the gold standard, The sensitivity of IFA for the detection of Leptospirosis was 38.8%, the specificity was 88.33%, the positive predictive value was 78.78%, and the negative predictive value was 56.38%. It seems that Leptospifrosis is widespread in Mazandaran province and the authorities should pursue prevention and control measures of this multifaceted disease with more enthusiasm to prevent spread of the disease. Demographic analysis of the results indicates that leptospirosis is typically an occupational disease in the region.

Background & Objective: Toxoplasmosis is a common infection among humans and animals. The disease is caused by Toxoplasma gondii. The prevalence of this infection depends on the weather conditions, dietary habits, and exposure to cats. This infection may lead to abortion, preterm labor and congenital anomalies in pregnant women. The present study is a descriptive cross-sectional study, which has been performed in order to determine the IgG antibody against toxoplasma in high-school girl students.Materials & Methods: In this study, 260 blood samples were randomly collected from the girl students in the high schools of Ilam in the year 2012. They were tested using the INDIRECT IMMUNOFLUORESCENCE method. In this study, a questionnaire was developed and presented to the considered population and the information like previous histories of eating underdone and roasted meat, exposure to cats, and place of residency were collected. In addition, the data were analyzed by SPSS software and chi-square test.Results: The prevalence rate of IgG antibody among these students was 21.1%. Chi-square test showed that there is a significant relationship between infection and eating underdone and roasted meat (P>0.002).Conclusions: Considering the low levels of anti-Toxoplasma gondii antibodies in high school girls, and non-immunity in pregnancy and the parasites’ irreparable effects on the fetus, it is essential to provide the necessary training before pregnancy to reduce the risks and problems.

History and Objectives: Strongyloides stercoralis infection has adverse effect on hemodialysis and renal transplanted patients. Since strongyloides stercoralis infection does not have apparent clinical signs, the present study was carned out to address the diagnostic power of ELISA and INDIRECT immunofluoresence on pateints referred to the hospitals for renal transplantation and hemodialysis in 1996.Materials and Methods: A diagnostic clinical trial was performed on 110 renal transplant and hemodialysis patients. ELISA, INDIRECT immunofluoresence, direct microscopic fecal examination and formalin-ether examination of fecal samples were compared. Sensitivity, specificity and predictive value of each method were determined.Results: From 10 samples, 14 showed infection whereas 96 samples did not show any infection. Diagnostic value was 100% for ELISA and INDIRECT immunofluoresent methods. There were close coorelation between the two mwthods. Both method of direct fecal examination formalin ether method were unable to detect all the positive samples and all negative samples were reported as such by two fecal examination methods. There were no significant difference on the detection of positive samples of hemodialysis patients of Gilan and Tehran province and between emodialysis and renal transplant patients of Gilan province. No significant difference between postive sample and sex was recorded.Conclusions: Immunosuppressant chemotherapy of rena l transplant patients causes reduction of inmmune response and low antibody titer and poor diognostic value of immunologic tests. ELISA and INDIRECT immunofluoresent detection of strongyloides stercoralis infection is very simple and rapid test to perform and has an high degree of sensitivity and specificity.

INDIRECT IMMUNOFLUORESCENCE (IIF) has been used to identify and measure autoantibody levels in pemphigus vulgaris but data about relationship between clinical severity of disease and antibody titers by IIF have been conflicting. We conducted this cross-sectional study to correlate the severity of oral and/or cutaneous involvement in patients with pemphigus vulgaris based on Pemphigus Area and Activity Score with IIF titers. Sixty-one new pemphigus vulgaris patients were included in this study. Human prepuce was used as substrate for IIF and assessment of disease severity was based on Pemphigus Area and Activity Score. The mean±SD age was 44.04±30.46 years, with a range of 18 to 79 years. IIF was positive in 56 (91.8%) patients. There was a significant relationship between total disease score and IIF titers ((P<0.001). Also a significant relationship was found between skin score (P=0.04) and mucosal score (P=0.04) with IIF titers. Our results show that there is a significant relationship between disease activity based on Pemphigus Area and Activity Score and antibody titers by IIF. Further studies are recommended to determine the usefulness of this technique for monitoring disease.

Aim and Background: Listeria monocytogenes is the ethiologic agent of abortion, preterm labour, neonatal septicemia and meningitis. The aim of this study was to determine the prevalence of listeriosis in aborted mothers or preterm labour. Two methods of INDIRECT immunofluorescens (IF) and amniotic culturing also were compared.Materials and Methods: This is an analytical study for evaluation of laboratory tests. In this research, the blood samples were collected from mothers and their serum were tested by INDIRECT IF for anti-listerial antibodies. Products of aborted fetus or preterm labour (amniotic fluid or a piece of placenta) were obtained as a specimen for culturing.Results: In this study, 518 mothers, 29 cases (5.59%) admitted with preterm labour, 125 cases (24.1%) with repeated abortion and 364 cases (70.31%) with first time abortion. Six cases (1.15%) had higher titer of anti-listeria antibody (1/800) in INDIRECT IF but in 5 cases (0.96%) L.monocytogenes was detected in culture.Conclusions: INDIRECT IF should be confirmed by culturing of infected specimens. Determining of anti-listerial antibody in pregnant women and selection of listeriosis for antimicrobial treatment may prevent the spontaneous abortion, still birth and preterm labour. Therefore, we suggest monitoring L.monocytogenes seropositivity in pregnant women with high risk of threatened abortion, and also microbiological assessment of symptomatic women for detection of L.monocytogenes and insidious infection.