ABSTRACT
Mycobacterium (M.) bovis, a bacterium in the M. tuberculosis complex, is a causative agent of bovine tuberculosis, a contagious disease of animals. Mycobacterial culture is the gold standard for diagnosing bovine tuberculosis, but this technique is laborious and time-consuming. In the present study, performance of the SD Bioline TB Ag MPT4 Rapid test, an immunochromatographic assay, was evaluated using reference bacterial strains and M. bovis field isolates collected from animals. The SD MPT64 Rapid test produced positive results for 95.5% (63/66) of the M. bovis isolates from cattle and 97.9% (46/47) of the isolates from deer. Additionally, the test had a sensitivity of 96.5% (95% CI, 91.2-99.0), specificity of 100% (95% CI, 96.7-100.0), positive predictive value of 100% (95% CI, 96.7-100.0), and negative predictive value of 92.9% (95% CI, 82.7-98.0) for M. bovis isolates. In conclusion, the SD MPT64 Rapid test is simple to use and may be useful for quickly confirming the presence of M. bovis in animals.
Subject(s)
Animals , Cattle , Cattle Diseases/diagnosis , Deer , Chromatography, Affinity/methods , Mycobacterium bovis/classification , Sensitivity and Specificity , Tuberculosis/diagnosisABSTRACT
PURPOSE: Tuberculosis (TB) is a major infectious disease and is responsible for two million deaths annually. For the identification and quantitation of Mycobacterium tuberculosis (M. tuberculosis), a causative agent of TB, a sandwich enzyme-linked immunosorbent assay (ELISA) against the MPT64 protein of M. tuberculosis, an antigen marker of the M. tuberculosis complex, was developed. MATERIALS AND METHODS: The MPT64 protein was expressed, and anti-MPT64 monoclonal antibodies were prepared. A sandwich ELISA was established using recombinant MPT64 protein and anti-MPT64 monoclonal antibodies. The sandwich MPT64 ELISA was evaluated using reference and clinical mycobacterial strains. RESULTS: The sandwich MPT64 ELISA detected MPT64 protein from 2.1 ng/mL to 250 ng/mL (equivalent to 1.7x10(4) CFU/mL and 2.0x10(6) CFU/mL). All 389 clinical M. tuberculosis isolates tested positive in the sandwich MPT64 ELISA (sensitivity, 100%), and the assay showed no cross reactivity to any tested nontuberculous mycobacterial strain (specificity, 100%). CONCLUSION: The sandwich MPT64 ELISA is a highly sensitive and quantitative test for MPT64 protein, which can identify M. tuberculosis.
Subject(s)
Antigens, Bacterial/analysis , Enzyme-Linked Immunosorbent Assay/methods , Mycobacterium tuberculosis/immunologyABSTRACT
Previously, we constructed a humanized antibody (HuS10) that binds to the common a antigenic determinant on the S protein of HBV. In this study, we evaluated its HBV-neutralizing activity in chimpanzees. A study chimpanzee was intravenously administered with a single dose of HuS10, followed by intravenous challenge with the adr subtype of HBV, while a control chimpanzee was only challenged with the virus. The result showed that the control chimpanzee was infected by the virus, and thus serum HBV surface antigen (HBsAg) became positive from the 14th to 20th week and actively acquired serum anti-HBc and anti-HBs antibodies appeared from the 19th and 23rd week, respectively. However, in the case of the study chimpanzee, serum HBsAg became positive from the 34th to 37th week, while actively acquired serum anti-HBc and anti-HBs antibodies appeared from the 37th and 40th week, respectively, indicating that HuS10 neutralized the virus in vivo and thus delayed the HBV infection. This novel humanized antibody will be useful in the immunoprophylaxis of HBV infection.
Subject(s)
Animals , Cricetinae , CHO Cells , Cricetulus , Hepatitis B/blood , Hepatitis B Antibodies/blood , Hepatitis B Surface Antigens/immunology , Hepatitis B virus/immunology , Neutralization Tests , Pan troglodytes/bloodABSTRACT
In a murine model system, enhancement of the total IgA and rotavirus-specific IgA of suckling mice was measured by ELISA with the intestinal fluid following oral administration of murine rotavirus EC (EDIM-Cambridge) strain. In the EC strain-administered group, the geometric mean titers (GMT) of total IgAs were 512 and 91 at 1 and 2 week postinfection, respectively. On the other hand, the GMTs of the rotavirus-specific IgAs were 108 and 3 at the same periods, respectively. Thus increase in the total IgAs was 64 folds and that in the rotavirus-specific IgAs was 43 folds compared with the negative control group. As the maximal titers of both the total and rotavirus-specific IgAs were observed at 1 week decreasing until 2 weeks after infection, it is evident that the GMT of the total IgA implies that of rotavirus-specific IgA. In our ELISA system, whose specificity was verified by Western blot analysis, the total IgA in the administered group was determined to be 40-400 ng per 1 ml of the intestinal fluid. Therefore it is concluded that determination of the rotavirus-specific IgA in murine models can be a sensitive indication of rotavirus infection, and will be another promising tool in viral challenge experiments in vaccine development.
Subject(s)
Animals , Mice , Administration, Oral , Blotting, Western , Enzyme-Linked Immunosorbent Assay , Hand , Immunoglobulin A , Mice, Inbred ICR , Rotavirus Infections , Rotavirus , Sensitivity and SpecificityABSTRACT
In Sprague Dawley (SD) rats, antibodies against strains of Orinentia tsutsugamushi, Kato, Karp and Gilliam, were produced in order to investigate their longevity and cross-reactivities to their corresponding homologous and heterologous antigens. By immunofluorescence assay (IFA) of IgG and IgM, it was shown that the immunity to the homologous strains persisted at a higher level (longevity of at least 34 weeks with higher IFA titers). On the other hand, the immunity to the heterologous strains persisted at a lower level (longevity of 10 to 34 weeks with lower IFA titers). Since infection with one strain of O. tsutsugamushi does not preclude reinfection with other strains, understanding of the antigenic diversity of O. tsutsugamushi and duration of the immunity to both homologous and heterologous strain is very important in diagnosis of scrub typhus.
Subject(s)
Animals , Rats , Antibodies , Antigenic Variation , Antigens, Heterophile , Diagnosis , Fluorescent Antibody Technique , Hand , Immunoglobulin G , Immunoglobulin M , Longevity , Orientia tsutsugamushi , Rats, Sprague-Dawley , Scrub TyphusABSTRACT
We immunized sixty two healthy subjects with the five different viral titers (300, 500, 1000, 1500 and 2000 plaque forming unit; pfu) of the MAV/06 strain of live attenuated Varicella-zoster virus (VZV) in order to gain sufficient information on safety and immuogenicity as a vaccine strain. Humoral immunity of all vaccine recipients was tested by the fluorescent antibody to membrane antigen (FAMA) assay and Enzyme-linked immunosorbent assay (ELISA) for the quantitative detection of IgG antibody. We tested neutralized antibody in 62 subjects by plaque reduction neutralization test (PRNT50). All of thirty two subjects with initial seronegative response had antibody by FAMA method at four weeks after immunization with four different preparations of dosage. The geometric mean titers (GMTs) of VZV antibody to membrane antigen was 160.9 in 6 subjects with 1500 pfu group; 83.3 in 14 subjects with 1000 pfu group: 116.2 in 7 subjects with 500 pfu groups and 72.0 in 6 subjects with 300 pfu group. Thirty subjects who had VZV antibody at the time before immunization demonstrated elevated antibody titer by FAMA assay and PRNT50 test. Side reactions of the vaccination was not demonstrated in all cases.
Subject(s)
Chickenpox , Enzyme-Linked Immunosorbent Assay , Herpesvirus 3, Human , Immunity, Humoral , Immunization , Immunoglobulin G , Membranes , Neutralization Tests , VaccinationABSTRACT
No abstract available.
Subject(s)
Enzyme-Linked Immunosorbent Assay , Herpes Simplex , Immunoglobulin G , Immunoglobulin MABSTRACT
No abstract available.
Subject(s)
Animals , Humans , Cell Line , Guinea Pigs , Guinea , Hantaan virusABSTRACT
No abstract available.