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Transplacental acquisition of cytomegalovirus antibody. Observations with two different CMV strains.

Titrations of complement fixing antibody (c.f.a.) were done in paired sera of 127 women at delivery and cord blood of their babies using two different cytomegalovirus strains both of which had undergone many passages in WI-38 cells, With High strain 74% of the mother and 41% of the babies were negative at the serum dilution 1:4. With Towne strain the incidence of negative was 5% both among the mother and the newborns. Worth of mention is the study of correlation of the titers in the matched pairs of sera according to the strain. With High strain 23 of the babies had c.f.a. titers significantly higher than their mothers and 14 of them were from negative women. This situation could be suggestive of an active placental transfer of IgG globulins. No such disparities were found using Towne strain as antigen. 48 pairs of sera were then tested with both strains by indirect hemagglutination, indirect immunofluorescence and gel precipitin test. Results are discussed in relation to the possible differences in the antigenic composition of the two cytomegalovirus strains.

Adult↗

Evidence against the presence of circulating immune complexes in chronic inflammatory bowel disease.

Several investigators have reported the presence of circulating immune complexes in serum from patients with Crohn's disease and chronic ulcerative colitis. Because previous assays employed conditions which might have caused immunoglobulin aggregates to form in vitro, thus falsely suggesting the presence of immune complexes in vivo, we tested inflammatory bowel disease sera for immune complexes using four assays designed to minimize in vitro immunoglobulin aggregation. In three assays immune complexes were not detectable, while in a fourth, the Clq precipitin test, positive reactions occurred. These precipitin reactions did not have characteristics of immune complexes. Our data suggest that circulating immune complexes are either not present in patients with inflammatory bowel disease or that they occur infrequently or in low concentration.

Antigen-Antibody Complex↗

Studies on the feeding patterns of the sheep headfly Hydrotaea irritans (Diptera, Muscidae) in Great Britain.

Serum-precipitin tests carried out on the stomach contents of laboratory maintained and field-caught Hydrotaea irritans showed that a positive result could be obtained for at least seven to 10 days after feeding. Eye, nasal and salivary secretions, even if undiluted, did not give a precipitin response in these tests. Field results from over 3000 flies thus indicate that 20 to 55% of the active fly population had taken a blood meal within the previous 10 days: 75 to 100% of these had fed on cattle blood and 15% or less on sheep blood. Significantly lower feeding levels were obtained from flies collected within plantations, indicating a protracted resting phase in woodland. From "fly feeding areas" high blood-feeding levels were shown from early July, as ovarian development begins. Early feeding rates were similar in both sexes.

Animals↗

Evaluation of alkaline phosphatase immunoassay and comparison with other diagnostic methods in areas of low transmission of schistosomiasis.

The alkaline phosphatase immunoassay (APIA) is an antibody detection technique which permits the diagnosis of schistosomiasis using a butanolic extract preparation from adult worms. APIA has demonstrated high sensitivity and specificity in previous reports with well characterized human sera. Its potential as a diagnostic tool for epidemiological surveillance was assessed in comparison with three other diagnostic tests: stool examination, ELISA with soluble egg antigen (SEA) and the circumoval precipitin test (COPT). APIA was 100% specific in an area without Schistosoma mansoni transmission and had 89% sensitivity in an endemic area where 69% of the infected subjects excreted less than 100 eggs g of faeces. It was found to be less sensitive in children under 5 years of age who were positive by the COPT test. APIA can be applied as an initial screening test, based on its high sensitivity, specificity, absence of cross-reactivity with intestinal parasites and the fact that it is a technique suitable for use in epidemiological surveillance.

Adolescent↗

Invasive aspergillosis in cancer.

Ten confirmed cases of invasive aspergillosis (IA) in cancer patients were analysed retrospectively. Eight were pulmonary, one was sinonasal and one was cutaneous. The majority of patients had haematological malignancies (7), the remaining three were cases of solid tumours. Fever was present in all 10 cases. Cough and lung signs were present in all eight cases of invasive pulmonary aspergillosis. Haemoptysis was encountered in three of nine cases of pulmonary and sinonasal aspergillosis. Mortality was low (2%). While corticosteroids, antibiotics and anticancer chemotherapy/radiotherapy were factors predisposing the patients to IA, neutropenia was perhaps responsible for their mortality. Seven of the patients had other associated pathogens isolated in culture in addition to Aspergillus spp. Aspergillus fumigatus was the predominant species, followed by A. flavus, A. glaucus, A. nidulans and A. niger. Direct microscopic examination (in six of seven cases) and culture (six of seven cases) correlated well with radiographic and clinical findings in cases with lung involvement. Serology for anti-Aspergillus antibodies performed by gel diffusion precipitin test was positive in one case of sinonasal aspergillosis, wherein only one precipitin band was observed. Correlation of clinical symptoms, consistent radiographic findings and microbiological work-up (the latter including a triad of direct microscopy, culture and serology) are required to arrive at a diagnosis of IA, especially where histology cannot form the mainstay of diagnosis.

Adolescent↗

A comparison of three serological tests for the diagnosis of B. ovis infection in rams.

The complement fixation test (CFT), the enzyme labelled immunosorbent assay (ELISA) and the gel diffusion precipitin test (GD) were compared, for the diagnosis of Brucella ovis infection in rams. The sensitivities of the tests in 109 rams which were shedding B. ovis in their semen were: CFT 96.3%; ELISA 97.2%; GD 91.7%. The specificities of the tests in 141 rams from non-infected flocks were: CFI 99.3%; ELISA 98.6%; GD 100%. Predictive values of the three tests were measured in 285 rams from infected flocks. Thirty-eight percent of these rams were shedding B. ovis in their semen. Predictive values of positive tests were: CFT 75.5%; ELISA 66.7%; GD 72.5%. Predictive values of negative tests were: CFI 97.1%; ELISA 97.6%; GD 93.8%.

Journal Article↗

Longitudinal serological study of malaria in infants in the West African savanna. Comparisons in infants exposed to, or protected from, transmission from birth.

Two infant populations, the one exposed to intense malaria transmission and the other protected, were followed and compared by six serological tests. The IgG and IgM levels increased with age and were systematically, though only slightly, lower in the protected children. The results of three Plasmodium falciparum tests (precipitin, indirect fluorescent antibody (IFA) and indirect haemagglutination and one P. malariae test (IFA) were high at birth and decreased rapidly afterwards in both populations. In the unprotected population, this decrease was followed by an increase, closely associated with the parasitological findings, while in the protected population the decrease continued to very low levels.

Africa, Western↗

Avian reovirus antibody assay by indirect immunofluorescence using plastic microculture plates.

An indirect fluorescent antibody test was developed to detect serum antibody to avian reovirus strain WVU2937. This test employed small multiple well plastic plates (8 x 5.5 cm) which readily fitted into the standard mechanical stage mechanism of an incident light fluorescence microscope. The small wells of the plates required minimal (10 muL) volumes of reagents. In tests on 18 sera in which the indirect fluorescent antibody, agar gel precipitin and plaque reduction methods were compared sera which gave negative results in the agar gel precipitin test were sometimes positive in the indirect fluorescent antibody and plaque reduction test, but indirect fluorescent antibody titers were lower than plaque reduction test titers. No false positive reactions were detected in 46 sera from uninoculated specific pathogen free chicks of up to eight weeks of age.

Animals↗

Fractionation of Schistosoma japonicum soluble egg antigen glycoproteins by hydrophobic interaction chromatography.

We are currently studying the soluble egg antigens of Schistosoma japonicum in an attempt to determine which antigens are potent immunogens. Previously, we demonstrated by Ouchterlony immunodiffusion and inhibition of the circumoval precipitin test that the glycoprotein fraction of soluble egg antigens contains the antigens which are most immunogenic in natural infections. The soluble egg antigen glycoproteins have now been further fractionated via hydrophobic interaction chromatography on phenyl Sepharose. We found that there were at least two antigens involved in the circumoval precipitin reaction. Both the hydrophilic antigen which we call japonicum antigen glycoprotein II (JAG II) and a mixture of hydrophobic antigens (JAG III and the JAG IV complex) were capable of causing a 50% inhibition of the COP reaction around S. japonicum eggs. JAG II was not a major serological antigen of S. japonicum since it gave only a weak precipitin line upon Ouchterlony immunodiffusion analysis with pooled sera from Filipino patients with chronic S. japonicum infections. Hydrophobic interaction chromatography yielded preparations which were sufficiently pure for use in radioimmunoassays. By radioimmunoassay, the best antigens among the glycoproteins were moderately hydrophobic JAG III and the JAG IV complex. They had large amounts of antibody directed toward them in patients with schistosomiasis japonica and exhibited little reactivity with S. mansoni. The hydrophilic glycoproteins JAG I and II were poor immunogens and extensively cross-reacted with S. mansoni. This cross-reactivity means that diagnostic tests with crude soluble egg antigens would run the risk of potential false-negative results in patients with other trematode infections.

Animals↗

Preparation and antigenicity of M protein released from group A, type 1 streptococcal cell walls by phage-associated lysin.

The lysis of Group A type 1 streptococcal cell walls by phage-associated lysin has been described. In the preparation of lysin, a new semisynthetic non-protein media to support growth of the propagating strain of Group C streptococci was employed. Following lysis of the cell walls, the resulting digest was partially purified by ion-exchange chromatography and ammonium sulfate precipitation. In addition to M protein, the resulting preparation (called lysin M protein) contained the group-specific carbohydrate and the T protein-but did not contain antigens, detectable by precipitin tests, which cross-reacted with absorbed heterologous group or type antisera. Capillary precipitin reactions between the lysin M protein and type-specific antiserum did not occur in the presence of high ionic strength buffers; these buffers did not similarly affect precipitin reactions of acid M protein. Type 1 lysin M protein is shown to be a good antigen. A total of 1.5 mg. injected intradermally in saline produced bactericidal antibody in eight of nine rabbits; when injected in adjuvant in one rabbit, protective serum antibodies developed. Streptococci grown in sera from seven of nine rabbits immunized with lysin M protein demonstrated significantly longer chains than when grown in normal rabbit serum. Antibody as demonstrated by each of these three tests was shown to be type-specific. No local or systemic toxicity was noted following intradermal injection in rabbits of lysin M protein.

Animals↗

Distribution of serotypes of Nocardia asteroides from animal, human, and environmental sources.

The antigenic types of 129 isolates of Nocardia asteroides from diverse clinical, environmental, and geographic origins were determined. The majority of the isolates studied were of bovine (56) or human (44) origin; 11 were derived from six species of animals other than cattle, and 10 were isolated from environmental sources; the source of 8 strains could not be determined. Testing culture filtrate antigens against four standard reference sera in a gel diffusion precipitin test established the antigenic type of 95.3% of the isolates. After excluding strains that weighted the data because of common infection, the distribution of serotypes was examined according to the origin of the isolate. Type I was the most frequently encountered serotype (31.9%); types III (15.0%) and IV (20.4%) were also observed frequently, as was the antigenic mixture III + IV (14.2%). There was an apparent difference in frequency of type III and IV antigens among isolates of bovine and human origin; type III made up 20.0% of the bovine isolates and 13.6% of the human isolates, whereas type IV constituted 10.0% of bovine and 27.3% of human isolates.

Air Microbiology↗

Evaluation of the specificity of Pasteurella multocida somatic antigen-typing antisera prepared in chickens, using ribosome-lipopolysaccharide complexes as inocula.

Purified lipopolysaccharides (LPS) from 16 serotypes of Pasteurella multocida were complexed with Aspergillus fumigatus ribosomes. The complexes were used as inocula to prepare antisera, in chickens, for somatic antigen typing by the gel diffusion precipitin test (GDPT). Antisera made against 15 of 16 LPS reacted with their respective specific heat-stable antigens in the GDPT and homologous LPS in the passive hemagglutination test. Antisera could not be made against serotype 15 LPS. Correlation was not observed between intensity of the precipitin reaction in the GDPT and titer to homologous LPS in the passive hemagglutination test. Most antisera cross-reacted with other heat-stable antigens of other serotypes in the GDPT. Many of these cross-reactions were eliminated by dilution. Cross-reactions that occurred in the GDPT with antisera made against LPS of serotypes 2, 5, 7, and 8 could not be eliminated by dilution.

Animals↗

Bluetongue virus in bovine semen: viral isolation.

Vero cell cultures and embryonating chicken eggs were used for direct isolation of bluetongue virus from cattle blood and from semen samples. Cell culture and embryonating chicken eggs each were more effective than was the blood autograft inoculation of susceptible sheep with selected blood and semen samples. Evaluation of the cell culture technique indicated that the quality of the distilled water was the primary factor responsible for the increased sensitivity of the Vero cell cultures for the present blue-tongue viral isolations. Test results showed that urine was a poor specimen for viral isolation when assayed in chicken eggs. A comparison of tests for precipitating and complement-fixing antibodies to bluetongue virus indicated that the precipitin test was the more accurate of the two tests.

Animals↗

Opportunistic endocarditis following open-heart surgery.

The incidence of endocarditis following open-heart operations is declining, but the relative incidence of endocarditis by the opportunistic organisms, both bacterial and fungal, has increased. These "opportunists" require careful identification and sensitivity testing so that appropriate therapy can be planned. In cases of candida endocarditis, a positive precipitin test and elevated candida antibody titer may provide an earlier means of diagnosis than blood cultures. These patients should be treated promptly with adequate dosages of specific antimicrobial or antifungal agents. Reoperation should be considered early if there is evidence of valve dehiscence or embolic phenomena or if drug therapy fails to control the infection.

Adult↗

The HemeSelect test: a simple and sensitive forensic species test.

The evaluation of a commercially available, faecal occult blood test to determine the species of origin of unknown stains is described. This test was simple to perform, requires a minimum amount of equipment and produced results that were easy to interpret. The HemeSelect test was found to be at least two orders of magnitude more sensitive than a precipitin test and would be the test of choice for small or aged stains.

Animals↗

Laboratory diagnosis of Schistosomiasis in areas of low transmission: a review of a line of research.

After 57 years of successful control of schistosomiasis in Venezuela, the prevalence and intensity of infection have declined. Approximately 80% of the individuals eliminate less than 100 eggs/g of stools, therefore morbidity is mild and the majority are asymptomatic. The sensitivity of Kato-Katz decreases to approximately 60%. Available serological methods for the detection of circulating antigens only reach a 70% of sensitivity. Tests based on the detection of antibodies by immunoenzymatic assays have been improved. The circumoval precipitine test has shown a high sensitivity (97%), specificity (100%), and correlation with oviposition, being considered the best confirmatory diagnostic test. Additionally to the classical immunoenzymatic assays, the development of the alkaline phosphatase immunoassay, allowed to reach a 100% specificity with an 89% sensitivity. Recently, we have developed a modified ELISA in which the soluble egg antigen is treated with sodium metaperiodate (SMP-ELISA) in order to eliminate the glycosilated epitopes responsible for the false positive reactions. The specificity and sensitivity reaches 97% and 99%, respectively. Synthetic peptides from the excretory-secretory enzymes, cathepsin B (Sm31) legumain (Sm32) and cathepsin D (Sm45), have been synthesized. The combination of two peptides derived from the Sm31 have been evaluated, reaching a sensitivity of 96% when analyzed independently and with a 100% specificity. Antibodies raised in rabbits against peptides derived from the Sm31 and Sm32 are currently evaluated in two different antigen-capture-based assays. The development of a simple, cheap and reliable test that correlates with parasite activity is a major goal.

Animals↗

Counterimmunoelectrophoresis (immunoelectroosmosis) and serum electrophoretic pattern in serologic diagnosis of canine blastomycosis.

Counterimmunoelectrophoresis (CIEP) with blastomyces and histoplasma antigens was used in a serologic study of 181 dogs clinically suspected of having blastomycosis and of 8 dogs with confirmed blastomycosis or histoplasmosis. Thirteen of the 181 dogs, positive by CIEP, were euthanatized, and the diagnosis was confirmed by cultivation and/or microscopic detection of Blastomyces dermatitidis. Additional CIEP-positive dogs were confirmed by staining of aspirates collected in vivo. Radiographic support for the diagnosis was reported in 4 other dogs in which histoplasmosis was excluded by a negative CIEP with histoplasma antigen. The precipitating antibody may disappear during the course of the disease, as it did in 1 dog treated with amphotericin B, but not cured. This dog reverted from CIEP-positive to CIEP-negative within 17 months of treatment (with a weak reaction after 10 months of treatment). The CIEP-detectable antibody was present only in 1 dog without a confirmation by histopathologic findings or cultivation among 24 well-documented cases and 181 total tested sera. The CIEP was more sensitive and specific than was the gel-diffusion precipitin test, eliminated the problems of anticomplementarity that often affected the results of complement-fixation tests with canine sera, and served well in detecting dogs with blastomycosis. Electrophoretic pattern of sera from CIEP-positive dogs with blastomycosis showed a decrease in albumin and an increase in alpha 2- and often in beta- and gamma-globulins, with a substantial decrease of the albumin/globulin ratio.

Animals↗