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[Detection of antigen of rickettsia of the tick group in bird droppings by antibody neutralization test].

The author presents methodical materials pointing to a possibility of using a micromethod of the antibody neutralization test for detection in bird pellets of the antigen of rickettsia referred to the Rocky Mountains spotted fever tick group. Specificity of the test, rickettsia antigen and sera was studied. The method is recommended for detection at the territory under study of circulation of the causative agent of tick-borne rickettsiosis by revealing the specific antigen in bird pellets.

Animals

Serotyping of avian infectious bronchitis viruses by the virus-neutralization test.

The antigenic relationships of 12 strains of infectious bronchitis virus (IBV) were evaluated by a virus-neutralization procedure similar to that used in typing human rhinoviruses. Such a procedure consists of reciprocal neutralization tests performed by reacting 32-320 EID50 or plaque-forming units of virus with 20 antibody units of antiserum. Eight serologic groups were identified by chicken embryo assay, and 4 by plaque-reduction (90%). In general, serologic groupings were not distinct but reflected numerous intergroup relationships. The contrasting results exhibited by indicator systems are viewed as differences in the accuracy of the methods employed. It is suggested that before an IBV classification scheme can be proposed, agreement must be reached on the most suitable indicator system, techniques must be standardized, and reference viruses and antisera distributed to several institutions for comparison testing. In addition, cross-protection studies are needed to determine the importance of IBV serotypes and/or variants on vaccine efficacy.

Animals

Antigenic relationships between flaviviruses as determined by cross-neutralization tests with polyclonal antisera.

The recently established virus family Flaviviridae contains at least 68 recognized members. Sixty-six of these viruses were tested by cross-neutralization in cell cultures. Flaviviruses were separated into eight complexes [tick-borne encephalitis (12 viruses), Rio Bravo (six), Japanese encephalitis (10), Tyuleniy (three), Ntaya (five), Uganda S (four), dengue (four) and Modoc (five)] containing 49 viruses; 17 other viruses were not sufficiently related to warrant inclusion in any of these complexes.

Antigens, Viral

Neutralization tests for dengue and Japanese encephalitis viruses by the focus reduction method using peroxidase-anti-peroxidase staining.

Neutralization tests were made on 4 types of dengue (DEN) virus and Japanese encephalitis (JE) virus by incubation of serially diluted antisera and constant amounts of the viruses and then focus assay of surviving virus infectivity with peroxidase-anti-peroxidase (PAP) staining. Neutralization reactions were virtually completed in 2 hr on incubation of serum-virus mixtures at 28 C. A straight regression line was obtained on a probit chart by plotting the focus reduction rates at various dilutions of a given serum against the logarithm of the serum dilution used in the test. The slopes of the probit regression lines for the neutralization for DEN types 1 and 3 were similar, but differed somewhat from those for DEN type 2 and type 4. The slope of the line for JE virus was quite different from those for DEN viruses. Using these relations, the fifty percent focus reduction titer (FR50) of neutralizing antibodies of a given serum could be estimated from the focus reduction rates at several dilutions of the test serum when the latter was between 25-75% of the value of the control.

Animals

The titration of tetanus antitoxin IV. Studies on the sensitivity and reproducibility of the toxin neutralization test.

The quality of tetanus toxin affected the sensitivity of the toxin neutralization (TN) test greatly. By using purified toxin a minimum level of 0.001 IU/ml of tetanus antitoxin could be detected whereas with crude toxin a level of 0.025 IU/ml only could be detected. The TN test described in this report permitted titration of tetanus antitoxin in twofold dilution steps from levels as low as 0.001 IU/ml using 0.6 ml of serum only at the L+/5000 level of purified tetanus toxin. Treatment of the sera with 2-mercaptoethanol (2-ME) did not affect the TN titres showing that the TN test detects the neutralizing antibodies (IgG) which are not affected by 2-ME. The TN test was found to be a highly sensitive and reproducible test.

Animals

A new lupus anticoagulant neutralization test based on platelet-derived vesicles.

We here present an easily standardizable and reproducible procedure which clearly separates lupus anticoagulants (LA) from coagulation factor inhibitors. This new LA neutralization test makes use of platelet-derived microvesicles which were prepared as follows: gel-filtered platelets (4 x 10(5)/microliters) were incubated with 60 microM of the calcium ionophore A23187 for 20 min at 37 degrees C. The vesicles were separated from the platelet aggregates by centrifugation at 1000 g for 10 min. The vesicle containing supernatant was then spun down at 15,000 g for 15 min, lyophilized and stored at -20 degrees C until used. The vesicles were resuspended in plasma from normal individuals, from patients with LA activity, from patients with factor VIII inhibitors, from patients with congenital factor deficiencies and from patients receiving oral anticoagulants or intravenous heparin. A kaolin clotting time was performed in the absence (KCT) or presence of these vesicles (KCTves) and the ratios of these times to their respective mean normal times were calculated. Segregation of LA patients from all remaining patients except heparinized ones could be made with a high degree of accuracy. A thrombin time was needed to separate LA from heparinized patients. The method was highly reproducible and only minor (negligible) differences in potencies were observed between different vesicle preparations. Both the intra-batch and the inter-batch coefficients of variations on the KCTves were lower than 6%.

Blood Coagulation Factors

Immunization against neonatal tetanus in New Guinea. 3. The toxin-neutralization test and the response of guinea-pigs to the toxoids as used in the immunization schedules in New Guinea.

The mouse toxin-neutralization test procedure described has been used for antitoxin titration of sera of New Guinea women following primary immunization with plain and adjuvant tetanus toxoids and following booster immunization, and for titration of guinea-pig sera.The responses of guinea-pigs immunized with the same toxoids and immunization schedule to simulate the human field trial studies were observed for 6 months. The relative antitoxin responses of guinea-pigs to the different toxoids were similar to those of the women.

Animals

A neutralization test survey for Lassa Fever activity in Lassa, Nigeria.

5-8% of 434 sera collected in Lassa village, Nigeria, in August 1970 were positive by neutralization test for Lassa fever (LF) virus. A second survey in March 1971 found 15 of 47 complete compounds tested in Lassa, Dille and Yuba villages had at least one peron with serologically demonstrable experience with LF virus. 67% of positive compounds had more than one infected member. Four of 17 hospital staff sero-converted between August 1970 and March 1971 without symptoms, evidence that subclinical infections of LF occr.

Adolescent

Vaccinia neutralization test using a radioisotope assay for virus-induced enzyme activity.

Vaccinia virus induction of a metabolic activity in host cell cultures forms the basis of a new assay for neutralizing antibodies. A direct relationship between the amount of vaccinia virus infecting cell cultures and the induced incorporation of tritium-labeled thymidine into the acid-insoluble fraction of the cells provided an indicator system. A liquid scintillation spectrometer was used to determine radioactivity associated with cell materials, and it provided a method for partial automation of an immunological procedure. Reproducibility of the method was satisfactory, and agreement with conventional vaccinia serum neutralization tests was demonstrated.

Amino Acids

Correlation between the serum neutralization test and the indirect immunofluorescent test for the detection of specific antibodies to infectious bovine rhinotracheitis virus.

Comparative titrations show that immunofluorescence on infected rabbit kidney cells is relatively as sensitive as the neutralization test for the detection of antibodies to the infectious bovine rhinotracheitis virus. When calf testis cells infected with this virus are used for the detection of antibodies by the indirect immunofluorescent test nonspecific staining is high. This nonspecific reaction is absent when rabbit kidney cells are used.

Animals

Semi-micro plaque neutralization test for rubella antibody.

Attenuated rubella virus, strain Cendehill, was found to produce small, distinct and ring-shaped plaques in Vero cell cultures. This finding was utilized to develop a semi-micro plaque neutralization test for a rubella antibody in 14 cmX9cm (24 wells) plastic trays. The procedure represents a reproducible and economical way to measure rubella neutralizing antibody.

Adult

[Local reaction and neutralization test in smallpox vaccination after previous inoculation with MVA. Stickl's graduated vaccination (author's transl)].

Of 200 first inoculations and 8 revaccinations, the sera were examined for content of neutralizing vaccine antibodies 3 weeks after the MVA graduated inoculation. In spite of imponderables which affect the failure rate in the "clinical trial", a positive result was obtained on first vaccination in 85%. In vaccinated children with a negative neutralization test a distinct booster effect was obtained with a "main vaccination" after about six months. The same effect was seen with revaccinations. For the time being inadequately controlled cerebral convulsive diseases, severe endogenous eczema, neurodermatitis, acute allergies, malignancies, immunosuppressive therapy, cortisole and intensive antirheumatic therapy and acute and purulent diseases must be considered as contraindications.

Adult