Twenty-four monoclonal antibodies were produced by immunizing BALB/c mice with Rickettsia tsutsugamushi Boryong strain and used for the analysis of antigenic characteristics of R.tsutsugamushi Boryong strain and antigenic heterogeneity of R.tsutsugamushi by indirect immunofluorescent(IF) test. R. tsutsugamushi Kato, Karp, Gilliam, TA686, TA716, TA763, TC586, TH1817, and Boryong were used for the analysis of antigenic heterogeneity of R.tsutsugamushi. Five monoclonal antibodies were reactive with 27-kDa protein, four monoclonal antibodies were reactive with 47-kDa protein, and eight monoclonal antibodies were reactive with 56-kDa protein of R.tsutsugamushi Boryong strain. The reactive protein of seven monoclonal antibodies could not be identified by immunoblotting method. All monoclonal antibodies to 27-kDa protein and three monoclonal antibodies to 47-kDa protein, and five monoclonal antibodies to 56-kDa protein were reactive with three to eight strains among nine strains of R. tsutsugamushi tested. One monoclonal antibody reactive to 47-kDa protein(KI18) and two monoclonal antibodies reactive to 56-kDa protein(KI36, and KI37) reacted with all the strains of R. tsutsugamushi tested. Strain-specific monoclonal antibody(KI58) could be found among antibodies which were reactive with 56-kDa protein. There was no strain which showed same reactivity pattern to these 24 monoclonal antibodies among nine strains. From this results, it could be concluded that Boryong strain is antigenically different from other strains of R.tsutsugamushi and antigenic heterogeneity of R.tsutsugamushi is due to the antigenic diversity of several proteins of R. tsutsugamushi including 56-kDa protein.
Animals ; Antibodies, Monoclonal/*immunology ; Antigens, Bacterial/*analysis ; Bacterial Proteins/analysis ; Mice ; Mice, Inbred BALB C ; Orientia tsutsugamushi/*immunology ; Species Specificity

OBJECTIVETo study the role of Mycoplasma pneumoniae (MP) load and antibody measurements in the diagnosis of MP pneumonia.

METHODSA total of 115 children with MP pneumonia and 400 healthy children were enrolled. The MP load and total antibody level were measured at different stages, and the MP load index (MPLI) was calculated.

RESULTSThe cut-off value of MPLI for MP infection was 6.12. MPLI and total antibody titer increased during the course of the disease, while MP-DNA decreased rapidly. Within the same time of blood collection, the group with a higher MP load had a significantly higher total antibody titer than the group with a lower MP load (P<0.05). Within 2 weeks of the course of the disease, the negative antibody group had a significantly higher MPLI than the positive antibody group (P<0.05).

CONCLUSIONSMPLI provides a standardized quantitative value of MP-DNA and plays an important role in the early diagnosis of MP infection.

Antibodies, Bacterial ; blood ; Child ; Child, Preschool ; DNA, Bacterial ; analysis ; Early Diagnosis ; Female ; Humans ; Infant ; Male ; Pneumonia, Mycoplasma ; diagnosis ; microbiology

Serum specimens from leprosy patients, their contacts and controls were examined for the presence of phenolic glycolipid I (PGL-I), a Mycobacterium leprae specific antigen, and antibodies to the antigen using enzyme-linked immunosorbent assays. Of 12 lepromatous patients with less than 2 years of therapy, 11(91.7%) were seropositive to PGL-l, thus indicating that new lepromatous cases can be identified by detecting anti-PGL-l antibodies. In contrast 88(56.4%) of 156 lepromatous patiens treated more than 2 years were positve. Moreover, only 69(40.8%) were seropositve among 169 lepromatous patients in the leprosy resettlement villages. The mean antibody level also declined significantly in proportion to the duration of chemotherapy. This may suggest the possibility of monitoring chemotherapy by detecting anti-PGL-l antibodies. The prevalence of anti-PGL-l antibodies among 200 controls from a high endemic area for leprosy was 5.5% and was significantly higher than that(1.5%) among 200 controls from a low endemic area. Of 103 household contacts in the resettlement villages, 10(9.7%) were seropositive, reflecting the frequent chance of exposure to M. leprae. However, PGL-l was not detected many in any of the sera from controls, contacts, and inactive lepromatous patients having the anti-PGL-l antibodies; on the other hand, 6(50%) of 12 lepromatous patients treated less than 2 years had detectable PGL-l in their sera. The results thus indicate that PGL-l detection may be more suitable for monitoring the effectiveness of chemotherapy and that it may be necessary to examine for the presence of PGL-l in sera from contacts and normal populations for confirming M. leprae infection.
Antibodies, Bacterial/*analysis ; Glycolipids/*blood ; Human ; Leprosy/*blood/diagnosis ; Serologic Tests ; Support, Non-U.S. Gov't

The changes of serum IgG antibody reactivity to protein antigens of Treponema pallidum after treatment of syphilis were observed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. Until 9 to 12 months after treatment, it was seen that there was a loss of several antibodies and some diminution in their reactivity in primary, secondary and early latent syphilis, but no changes occurred in late latent and reinfected syphilis. In primary syphilis, there was a significant loss of two IgG antibodies to the treponemal antigens of molecular weights 68,500 and 47,000 at 11 months after treatment. According to our previous study, the treponemal antigen of molecular weight 68,500 was T. pallidum specific and appeared only in primary syphilis, and that of molecular weight 47,000 was one of the major antigens of T. pallidum. The reaction between serum IgG antibodies of 14 patients who had been treated for secondary, early latent and late latent syphilis 2 to 14 years ago and major antigens of T. pallidum was observed and any loss or decrease in reactivity was not discovered. From the results obtained, it was concluded that the observation of serum IgG antibody reactivity to protein antigens of T. pallidum is not helpful in evaluating the efficacy of treatment in secondary, early latent, late latent and reinfected syphilis. However, serum IgG antibodies to treponemal antigens of molecular weights 68,500 and 47,000 could possibly be useful in the assessment of the efficacy of treatment in primary syphilis.
Antibodies, Bacterial/*immunology ; Antigens, Bacterial/*analysis ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Humans ; Immunoglobulin G/*analysis ; Recurrence ; Syphilis/*diagnosis/immunology/therapy ; Time Factors ; Treponema pallidum/*immunology

Five monoclonal antibodies (MAbs) and chicken immunoglobulin (IgY) were developed by immunizing with flagella purified from Listeria monocytogenes 4b and the five MAbs have been confirmed to be specific against three different epitopes of flagellin. The antibodies showed specific reaction to Listeria genus and no cross-reactivity with other bacteria tested in this experiment including E.coli O157:H7 and Salmonella enteritidis. Sandwich enzyme-linked immunosorbent assays (ELISA) using the MAbs and IgY were developed to detect Listeria species and the sensitivity and specificity of the developed ELISA have been analyzed. The detection limit of ELISA using MAb 2B1 and HRP labeled IgY was 1 x105cells/0.1 ml at 22degrees C and 1x106 cells/0.1 ml at 30degrees C. ELISA using the pair of MAbs (MAbs 2B1 and HRP labeled MAbs 7A3) detected up to 104cells/0.1 ml at 22degrees C and 30degrees C. Detection limit of sandwich ELISA using IgY was 10 times lower than MAb pair. Using the developed ELISA, we could detect several Listeria contaminated in food samples after 48 h-culturing. In conclusion, both MAbs and IgY have been proved to be highly specific to detect Listeria flagella and the developed sandwich ELISA using these antibodies would be useful tool for screening Listeria spp. in food.
Animals ; Antibodies, Bacterial/*chemistry ; Antibodies, Monoclonal ; Antibody Specificity ; Antigens, Bacterial/analysis ; Enzyme-Linked Immunosorbent Assay/*methods ; Flagella/*genetics ; Food Microbiology ; Immunoglobulins/analysis ; Listeria/*classification/immunology/*isolation&purification ; Meat/microbiology ; Milk/microbiology ; Sensitivity and Specificity ; Swine

There are many serologic tests for syphilis. By means of the usual serologic tests, it is not possible to differentiate between patients who need therapy and those who are cured. In this paper I want to discuss the scientific developments and demonstrate the results of immunologic research in syphilis, which makes it possible to differentiate between treated and untreated cases.
Antibodies, Bacterial/analysis ; Chromatography, Gel ; Electrophoresis, Starch Gel ; Fluorescent Antibody Technique ; Hemagglutination Tests ; Human ; Immunoglobulin G/analysis ; Immunoglobulin M/analysis ; Syphilis/immunology* ; Syphilis Serodiagnosis* ; Treponema pallidum/immunology

We utilized polyethylenimine (PEI) and Glu solution to immobilize the antibody of Treponema Pallidum on the silver electrode of piezoelectric quartz crystal and detect the standard antigen solution at different concentration. The antibody keeps the intrinsic Y type molecular structure revealed by AFM. The resonant range of the sensor is between 5 x 10(-5) - 1.25 x 10(-4) g/mL and the correlation coefficient is 0.9976, the optimal resonance pH is 7.5. We found the sensor having good selectivity in comparison with the method using BSA. At last, a discussion on the reproducibility of the sensor is presented.
Antibodies, Bacterial ; immunology ; Antigens, Bacterial ; analysis ; Biosensing Techniques ; instrumentation ; Equipment Design ; Immunoassay ; instrumentation ; methods ; Microelectrodes ; Polyethyleneimine ; Quartz ; Treponema pallidum ; immunology ; isolation & purification

Monoclonal antibody against kanamycin was prepared, and competitive direct ELISA and immunochromatographic assay were developed using the antibody to detect kanamycin in animal plasma and milk. The monoclonal antibody produced was identified to be IgG1, which has a kappa light chain. No cross-reactivity of the antibody was detected with other aminoglycosides, indicating that the monoclonal antibody was highly specific for kanamycin. Based on competitive direct ELISA, the detection limits of kanamycin were determined to be 1.1 ng/ml in PBS, 1.4 ng/ml in plasma, and 1.0 ng/ml in milk. The concentration of intramuscularly injected kanamycin was successfully monitored in rabbit plasma with competitive direct ELISA. Based on the colloidal gold-based immunochromatographic assay, the detection limits of kanamycin were estimated to be about 6-8 ng/ml in PBS, plasma, and milk. The immunochromatographic assay would be suitable for rapid and simple screening of kanamycin residues in veterinary medicine. Screened positives can be confirmed using a more sensitive laboratory method such as competitive direct ELISA. Therefore, the assays developed in this study could be used to complement each other as well as other laboratory findings. Moreover, instead of slaughtering the animals to obtain test samples, these methods could be applied to determine kanamycin concentration in the plasma of live animals.
Animals ; Anti-Bacterial Agents/*analysis ; Antibodies, Monoclonal ; Chromatography/methods/veterinary ; Enzyme-Linked Immunosorbent Assay/methods/*veterinary ; Kanamycin/*analysis ; Mice ; Milk/*chemistry ; Rabbits

OBJECTIVETo explore etiology, clinical manifestation and immunological changes of infectious pneumonia of neonates in Chengdu area.

METHODSSerum specimens were collected from 111 infants with infectious pneumonia. Eight viral and mycoplasmal specific serum IgM antibodies were detected by enzyme linked immunosorbent assay (ELISA); C reactive protein (CRP), total IgG and its subclasses, IgA and IgM were determined by rate scattered nephelometry; T lymphocyte subpopulations were detected by biotin-streptavidin-peroxidase method, and clinical and other laboratory data were analyzed.

RESULTS(1) Etiological agents: specific serum IgM antibodies were positive in 40 of 111 cases (36.0%) with pneumonias. All the 30 control infants were negative for the specific serum IgM antibodies. Among 111 infants with infectious pneumonia, 20.7% had single viral or mycoplasmal infection, 40.5% had bacterial infection, 15.3% had viral and mycoplasmal infection with bacterial infection; 23.4% had infection with unknown agents. (2) The most common clinical manifestations were tachypnea and cyanosis. The next were cough, milk choking, rales, retractions of the supraclavicular, intercostal and subcostal areas. Roentgenographic examination commonly revealed vague opacities, increased density and patchy infiltration. (3) Immune status: (1) CD(3), CD(4) cell counts of infants with pneumonias were lower than those of the controls while their serum IgA, IgM concentrations were higher than those of the control. (2) The CD(3) and CD(4) cell counts of the group with bacterial infection were lower than those of the control group. (3) The serum IgA concentration of the group with viral and mycoplasmal infection was higher than those of the control group and the group with unknown infection. (4) The serum IgM concentration of the group with bacterial infection was higher than those of the control group. (5) There were no significant differences in CD(8) cell counts, CD(4)/CD(8), concentration of serum IgG and IgG(1 - 4) between pneumonia group and the control group, and among various infectious groups and the control.

CONCLUSIONPathogens of neonatal infectious pneumonia in Chengdu area included single viral or mycoplasmic infection or bacterial infection, viral and mycoplasmal infection with bacterial infection, and unknown infection. Immunological changes of newborn infants suffered from infectious pneumonia included declined CD(3) and CD(4) cell counts, particularly in bacterial infection.

Antibodies, Bacterial ; blood ; Antibodies, Viral ; blood ; Bacterial Infections ; complications ; C-Reactive Protein ; analysis ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin M ; blood ; Infant, Newborn ; Male ; Pneumonia ; diagnosis ; etiology ; immunology ; T-Lymphocyte Subsets ; immunology ; metabolism ; Virus Diseases ; complications

The filtrate of Staphylococcus aureus culture has been used in an ampule form named as staphylococcal enterotoxin C injection for cancer therapy in clinic for ten years in China and proved to be effective. The active constituent of three kinds of injections is claimed to be staphylococcal enterotoxin C2 (SEC2), and the content of SEC2 is used as quality control. However, the correct content of SEC2 was not known and the relative amount of SEC2 was very low because of the complicated components of the filtrate. In this research, we established a proper ELISA system for the detection of SEC2 in staphylococcal enterotoxin C injection, which will improve the quality control of the injection. We produced and identified polyclonal and monoclonal antibodies of SEC2 and established BA-ELISA method based on the method of sandwich ELISA. It was found that the BA-ELISA method had good specificity, sensitivity and reproducibility, and being able to detect SEC2 at concentration from 2 to 20 ng x mL(-1), with an average CV value of 5.08%. The SEC2 content in staphylococcal enterotoxin C injection was calculated. There is some difference between the actual and labeled contents in the injections.
Animals ; Antibodies, Bacterial ; biosynthesis ; immunology ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antineoplastic Agents ; administration & dosage ; analysis ; immunology ; Enterotoxins ; administration & dosage ; analysis ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Hybridomas ; secretion ; Injections ; Mice ; Mice, Inbred BALB C ; Quality Control ; Rabbits ; Staphylococcus aureus ; chemistry