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Agglutination Test (agglutination + test)
Kinds of Agglutination Test Selected AbstractsSpecific Fab fragments recovered by phage display technique recognizing human spermatozoaINTERNATIONAL JOURNAL OF ANDROLOGY, Issue 5 2009Dorota Fiszer Summary Human hybridoma cell lines are often unstable and loose ability for antibody production. Sometimes, they show low and varying levels of heavy and light chains synthesis. Therefore it is reasonable to preserve generated specificities of light and heavy chains by cloning them to phagemid vector and creating phage display library. The aim of this study was to construct phage display library of Fab fragments recognizing sperm surface antigens. The source of mRNA constituted seven hybridoma cell lines producing antisperm antibodies which was proved by ELISA, and agglutination test as well as by inhibition of sperm to penetrate hamster oocytes. Fragments of cDNA encoding ,/, and , chains were cloned into pComb3HSS phagemid vector and amplified in XL-1Blue. The library was panned against whole unfixed sperm cells. Three positive clones selected after fourth round of panning showed heavy chain belonging to VH4 family, two of them (G28, K61) possessed lambda chain from VL2 family and one (H43) kappa chain from VK1 family. As these Fabs revealed similarities to antibodies against some proteins involved in sperm motility and cell fusion it can be suggested that these Fabs may be a cause of infertility. Finally, we proved that it is feasible to preserve specificities produced by human hybridomas using phage display technique and we recovered some Fabs which may be of diagnostic and research value, and may also have some value for contraceptive vaccine. [source] Cutaneous cryptococcosis associated with lepromatous leprosyINTERNATIONAL JOURNAL OF DERMATOLOGY, Issue 6 2001Rubem David Azulay MD A 65-year-old Brazilian man presented with an erythematous nodular lesion on the left forearm (Fig. 1). The patient had been treated with multidrug therapy for 8 months for lepromatous leprosy. During therapy, he developed recurrent episodes of reactions which were treated with high doses of prednisone and thalidomide. The histopathology of the cutaneous nodular lesion showed a granulomatous inflammatory infiltrate; some histiocytes contained vacuolations and others demonstrated oval-like or coma-like structures (Fig. 2). The specimen was cultivated in Sabouraud agar at room temperature. The colonies were transferred to Petri dishes containing Niger Seed Agar (NSA) (Fig. 3). The confirmed diagnosis was Cryptococcus neoformans var. neoformans based on microscopy and physiology, including the canavanine,glycine,bromothymol blue (CGB) medium (Lazéra MS, Pires FDA, Camillo-Coura L et al. Natural habitat of Cryptococcus neoformans var. neoformans in decaying wood forming hollows in living trees. J Med Vet Mycol 1996; 34: 127,131). The liquor culture was negative. Hemoculture and urine culture were also negative. Latex agglutination test was blood positive and liquor negative. Figure 1. Erythematous nodular lesion on the left forearm measuring 9 cm in diameter Figure 2. Granulomatous infiltrate presenting oval-like or coma-like structures inside the histiocytes (mucicarmine stain, ×,100) Figure 3. Petri dishes with Niger Seed Agar containing numerous colonies of Cryptococcus neoformans var. neoformans The patient's hemogram revealed normocytic anemia and normal total and differential white blood count. The CD4 count was 189/m3 and the CD8 count was 141/m3. Serology for anti-human immunodeficiency virus-I (anti-HIV-I) antibodies was negative. The X-ray of the lungs showed an areolar image in the superior lobe of the right lung. Therapy with prednisone was suspended and fluconazole (300 mg/day) was prescribed. The nodular cutaneous lesion regressed completely after 90 days. The patient was submitted to a second skin biopsy for treatment control. The culture of the specimen taken was still positive and the histopathology showed the same picture as before treatment. After 5 months of continued therapy with fluconazole, another biopsy was performed but no fungus was recovered from the specimen. [source] Clinicopathologic Features and Outcome Predictors of Leptospira interrogans Australis Serogroup Infection in Dogs: A Retrospective Study of 20 Cases (2001,2004)JOURNAL OF VETERINARY INTERNAL MEDICINE, Issue 1 2007Cinzia Mastrorilli Background and Hypothesis: We retrospectively evaluated the Clinicopathologic findings and outcome predictors in dogs with Leptospira interrogans Australis serogroup infections. Animals and Methods: The medical records of 159 dogs that had a leptospiral microscopic agglutination test (MAT) performed between 2001 and 2004 were reviewed. Results: Twenty dogs met serologic criteria for either symptomatic (16 dogs) or asymptomatic (4 dogs) infection caused by Leptospira interrogans Australis serogroup. Seven of 16 symptomatic dogs died or were euthanized and 9/16 recovered. Systemic inflammatory response syndrome (SIRS) was observed in 9/16 dogs. The presence of SIRS did not affect prognosis (P= .357). C-reactive protein (CRP) and haptoglobin (Hpt) concentrations were altered in all symptomatic dogs, but results did not differ significantly between survivors and nonsurvivors (P= .08 and P= .055, respectively). Conversely, the CRP to Hpt ratio (CRP/Hpt) was significantly increased in nonsurvivors. Disseminated intravascular coagulation (DIC) was diagnosed in 7/16 dogs. DIC did not significantly affect outcome (P= .126). Multiple organ involvement was present with renal failure in 16/16, liver damage in 12/16, cardiac damage in 11/16, and muscular damage in 8/16 dogs. Conclusions and Clinical Importance: Among the evaluated Clinicopathologic biomarkers, serum albumin, cardiac troponin I, CRP/Hpt, urinary albumin, and urinary total protein to creatinine ratio were found to predict outcome and warrant evaluation in larger prospective studies. [source] Evaluation of a reversed passive latex agglutination test for the detection of Verocytotoxin (VT) expressed by strains of VT-producing Escherichia coliLETTERS IN APPLIED MICROBIOLOGY, Issue 6 2001H. Chart Aims: To compare an experimental Reversed-Passive Latex Agglutination (RPLA) with Vero cells for the detection of Verocytotoxin expressed by VT-producing strains of Escherichia coli (VTEC). Methods and Results: The RPLA was used alongside a Vero cell tissue culture assay for the detection of VT in bacterial culture supernatant fluids and patients' faecal extracts. Conclusions: The RPLA was comparable with the Vero cell assay, although slightly less sensitive. Significance and Impact of the Study: The RPLA test proved to be a simple, rapid and convenient method of detecting VT in bacterial culture supernatant fluids and in the faeces of patients infected with VTEC. [source] Application of immunoproteomics to leptospirosis: towards clinical diagnostics and vaccine discoveryPROTEOMICS - CLINICAL APPLICATIONS, Issue 4 2007Uraiwan Kositanont Abstract Each of the currently available methods for serodiagnosis of leptospirosis, including the microscopic agglutination test (MAT), has its own drawback(s) when used in clinical practice. A new diagnostic test is therefore required for an earlier and more accurate diagnosis of leptospirosis. We applied immunoproteomics to define potential immunogens from five serovars of Leptospira reference strains. A leptospiral whole cell lysate from each serovar was used as the antigen to react with IgG and IgM in the sera from four patients with a positive MAT. Sera from four non-leptospirosis patients with a negative MAT were pooled and used as the negative control. 2-D Western blot analysis showed that the degree of immunoreactivity corresponded with the MAT titers. No immunoreactive spots were detected when the pooled control sera were used. A total of 24 protein spots immunoreacted with IgM and/or IgG from patients with leptospirosis. These immunoreactive proteins were identified by MALDI-TOF MS and were classified into five groups, including flagellar proteins, chaperones/heat shock proteins, transport proteins, metabolic enzymes, and hypothetical proteins. More immunoreactive spots were detected with anti-human IgG in the sera of all patients and with all the serovars of leptospires used. Some of the identified proteins immunoreacted only with IgG, whereas the others were detectable with both IgM and IgG. Among the immunoreactive proteins identified, FlaB proteins (flagellin and flagellar core protein) have been shown to have a potential role in clinical diagnostics and vaccine development. These data underscore the significant impact of immunoproteomics in clinical applications. [source] Changes of Circulating Antibody Levels Induced by ABO Antibody Adsorption for ABO-Incompatible Kidney TransplantationAMERICAN JOURNAL OF TRANSPLANTATION, Issue 5 2009P. V. Valli ABO-incompatible kidney transplantation using immunoadsorption to remove anti-A/B antibodies has become a successful clinical practice. Since the data on the specificity of the ABO columns are controversial, the present study assessed the efficiency and specificity of the ABO immunoadsorption, the effect on total immunoglobulins and antibodies previously induced by vaccination. Anti-A/B antibodies were measured by agglutination and ABO flow cytometry, total IgG/IgM, carbohydrate- and protein-specific antibodies by nephelometry and ELISA. The first immunoadsorption not only efficiently reduced donor-specific anti-A/B IgM (81%) and IgG (56%) but also reduced compatible anti-A/B IgM (59%) and IgG (34%). The measurements of antidonor A/B antibodies by direct agglutination (IgM) or flow cytometry better represented the effective antibody levels than the indirect agglutination test (IgG). The median reduction of total IgM and total IgG levels after a single immunoadsorption was 34% and 18%, respectively. Antibodies against pneumococcus and haemophilus polysaccharide antigens were significantly reduced, whereas antitetanus and antidiphtheria protein antibodies were not affected. Intravenous immunoglobulin administration restored the protective anticarbohydrate antibody levels. In summary, immunoadsorption efficiently removed antidonor A/B antibodies, but was not specific for A/B antigens. Anti-A/B antibody levels as determined by ABO flow cytometry are useful to establish the minimal number of immunoadsorptions needed for successful ABO-incompatible transplantation. [source] Serum sperm antibodies unrelated to mumps orchitisANDROLOGIA, Issue 2 2001S. Kalaydjiev In order to determine whether there is an association between mumps orchitis and serum sperm antibodies, we tested patients at the time orchitis was diagnosed (n=7) and individuals who had had orchitis at least 1 month previously (n=14). Data were compared with the results for a control group of blood donors (n=20). Sperm antibodies were detected by the gelatin agglutination test of Kibrick, the tray agglutination test of Friberg and the ELISA. Clinically significant sperm antibody levels were not found in patients in the early stages of the disease. Four subjects tested post-disease were positive in the Friberg test and one was positive in the ELISA. One control serum was also positive in the latter test. Significant differences were not found between levels in patients in the early stages of the disease and levels in individuals post-disease, although the results of the Friberg test differed significantly between controls and former mumps orchitis cases. These data do not support the assumption of an involvement of humoral immunity against spermatozoa in mumps orchitis patients. [source] Performance of commercial latex agglutination tests for the differentiation of Candida dubliniensis and Candida albicans in routine diagnostics,APMIS, Issue 11 2007E. CHRYSSANTHOU Candida dubliniensis is phenotypically similar to Candida albicans and may therefore be underdiagnosed in the clinical microbiology laboratory. The performance of Bichro-Dubli latex agglutination test for rapid species identification of C. dubliniensis was prospectively evaluated on 111 vaginal and 118 respiratory isolates. These had presumptively been identified as C. albicans/C. dubliniensis by their green colonies on CHROMagar Candida plates. Bichro-Dubli test identifed 2 (1.8%) vaginal and 6 (5.1%) respiratory isolates as C. dubliniensis. The test was also positive for 37 C. dubliniensis control strains characterised by 18S-28S DNA-sequencing. Bichro-Dubli test is thus a sensitive and accurate tool for rapid diagnostics in routine laboratories. [source] A rapid latex agglutination test for gender identification in the Atlantic bluefin tuna, Thunnus thynnus (Linnaeus)AQUACULTURE RESEARCH, Issue 9 2010Elisabetta Micera Abstract A rapid, one-step agglutination assay has been developed, based on latex particles sensitized with antibodies against vitellogenin (Vtg), aimed at Atlantic bluefin tuna, Thunnus thynnus (Linnaeus) (ABFT), gender identification. The egg-yolk precursor protein Vtg was used as a gender marker for the assay as it is a female-specific protein synthesized during reproductive maturation. The presence of Vtg in the plasma was revealed in 60,120 s through an agglutination reaction by mixing small volumes of ABFT plasma and an anti-Vtg antibody-latex suspension on a microscope slide. The effectiveness of the present test was restricted to the months of May and June, concomitant with high circulating Vtg levels. Because of its rapidity and ease of performance in the field, the present gender identification assay could be useful for broodstock management in the aquaculture industry as well as in tagging studies on wild populations. [source] Prevalence of antibodies to Leptospira serovars in beef cattle in central QueenslandAUSTRALIAN VETERINARY JOURNAL, Issue 5 2001PF BLACK Objective To obtain up-to-date data on the prevalence of antibodies to Leptospiraserovars in central Queensland beef herds preliminary to assessing their role in bovine subfertility and the role of cattle as a zoonotic reservoir. Design Sera from 2857 female cattle in 68 central Queensland beef herds were tested for antibodies to 14 Leptospiraserovars using the microscopic agglutination test. Vaccination use and age of cattle were collected to enable the calculation of crude and age-stratified seroprevalences. Results The most commonly detected antibodies were to serovars hardjo (15.8% crude seroprevalence), tarassovi (13.9%), pomona (4.0%) and szwajizak (2.2%). Vaccinates were omitted from the hardjo and pomona seroprevalence data. The seroprevalence for hardjo and pomona tended to increase with age of the animals. Conclusion These results are broadly similar to those of previous serological surveys. The data suggest that serovars other than hardjo, pomona and tarassovi, are unlikely to have a significant role in bovine subfertility and that cattle are unlikely to be a source of human infection with them in central Queensland. [source] Surveys in Papua New Guinea to detect the presence of Trypanosoma evansi infectionAUSTRALIAN VETERINARY JOURNAL, Issue 12 2000SA REID Objective To confirm serological evidence that Trypano-soma evansi is present in Papua New Guinea. Design Three surveys were undertaken in PNG during 1997/1998. Animals were selected for sampling on the basis of convenience. Samples of blood were examined for the presence of T evansi b y the haematocrit centrifugation technique (HCT) and mouse inoculation test (MI). Sera were tested in the field using the card agglutination test for trypanosomiasis/T evansi(CATT). Bovine sera were tested at James Cook University using an antibody-detection ELISA (Ab-ELISA). Results from testing bovine sera with the Ab-ELISA and sera from wallabies with the CATT were analysed using FreeCalc to determine the probability that animals in these populations were infected with T evansi. Results A total of 545 serum samples were collected. during the three surveys of which 39 cattle, two pig and three agile wallaby samples were positive with the CATT. All bovine sera collected were negative when tested with an Ab-ELISA. T evansi was not isolated using the HCT or the MI from any of these animals. Conclusion Based on the Ab-ELISA results it was concluded that T evansi infection was not present in cattle in villages around Balimo at a minimum expected prevalence of 10% (P < 0.05) and, based on the CATT results, that infection was not present in wallabies on the Bula plain at a minimum expected prevalence of 10% (P < 0.1). These results indicate that it is unlikely that T evansi is endemic in PNG [source] Identification and serotyping of atypical Legionella pneumophila strains isolated from human and environmental sourcesJOURNAL OF APPLIED MICROBIOLOGY, Issue 1 2007J.H. Helbig Abstract Aims:, To validate identification methods for Legionella pneumophila strains that cannot be serotyped into the known serogroups and to characterize their antigenic diversity. Methods and Results:, Fifty L. pneumophila strains that could not be serogrouped, but which had been confirmed as L. pneumophila by mip gene sequencing, were further identified phenotypically. We used (i) MONOFLUO anti- Legionella Staining Reagent (Bio-Rad) (50/50), (ii) an in-house prepared immunoblot assay for the detection of L. pneumophila - specific Mip protein epitope (50/50), (iii) fatty acid analysis using the Microbial Identifications System (MIDI) (47/50) and (iv) Oxoid agglutination tests (44/50). The serological diversity was further characterized by testing with five serogroup-cross-reactive monoclonal antibodies, resulting in nine phenons. Conclusions:, The division of L. pneumophila into 15 serogroups does not reflect the serogroup heterogeneity. Results of these tests indicate that there are more serogroups. Significance and Impact of the Study:, MONOFLUO anti- Legionella Staining Reagent is the only commercially available tool for identifying atypical strains of L. pneumophila. If necessary for epidemiological purposes, the antigenic heterogeneity of these strains can be analysed by monoclonal antibodies. [source] Performance of commercial latex agglutination tests for the differentiation of Candida dubliniensis and Candida albicans in routine diagnostics,APMIS, Issue 11 2007E. CHRYSSANTHOU Candida dubliniensis is phenotypically similar to Candida albicans and may therefore be underdiagnosed in the clinical microbiology laboratory. The performance of Bichro-Dubli latex agglutination test for rapid species identification of C. dubliniensis was prospectively evaluated on 111 vaginal and 118 respiratory isolates. These had presumptively been identified as C. albicans/C. dubliniensis by their green colonies on CHROMagar Candida plates. Bichro-Dubli test identifed 2 (1.8%) vaginal and 6 (5.1%) respiratory isolates as C. dubliniensis. The test was also positive for 37 C. dubliniensis control strains characterised by 18S-28S DNA-sequencing. Bichro-Dubli test is thus a sensitive and accurate tool for rapid diagnostics in routine laboratories. [source] | |||||||||||||||||||||||||