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PubMed1988

Immunization against experimental rabbit cysticercosis using liposome-associated antigen preparations.

Rabbits were vaccinated once, by subcutaneous and intradermal injection, with sonicates of oncospheres (TpO) or conditioned media from in vitro maintained mature metacestodes (TpMcES) of Taenia pisiformis. Extracts were either incorporated into or mixed with unilamellar liposomes (reverse phase evaporative vesicles) or emulsified in Freund's Incomplete Adjuvant (FIA). Control groups received liposomes or FIA without antigen, or antigen preparation without adjuvant. Rabbits were challenged orally two weeks after vaccination with approximately 1500 eggs of T. pisiformis and necropsied eight weeks after challenge. A mean of 155 cysts was recovered from seven control rabbits. A 67% reduction in peritoneal cyst numbers was obtained in TpO-IFA vaccinated rabbits compared to 75% for the TpO-liposome entrapped group. The highest level of protection (86%) was obtained when TpO was mixed with but not entrapped in liposomes. Only 32% and 39% reduction in peritoneal cyst numbers was obtained after immunizing with the TpMcES preparation in liposomes or IFA respectively, however greater than 85% of peritoneal metacestodes were dead (necrotic or calcified) and suggests a different immune response than occurs after vaccination with oncosphere extracts. Specific anti-oncospheral or anti-metacestode ES antibody (IgG) responses at two weeks post vaccination were similar in rabbits immunized with liposome or IFA associated extracts.

Animals↗
PubMed1975

Serologic evidence of the postoperative persistence of hydatid cysts in man.

The sensitivity of the haemagglutination, complement-fixation and indirect fluorescent antibody tests in the diagnosis of primary and recurrent hydatid disease is presented. In the first, diagnostic titres were obtained in 90% of 50 patients with hepatic and 64%, 60%, and 68% of 25 patients with pulmonary cysts, respectively. In the postoperative study of 62 patients, elevated titres of HA, CF and IFA antibodies in 30 led to the presumptive diagnosis of recurrent disease, confirmed later by radiological and surgical follow-ups. In the 32 others the proportion of positive reactors was 29%, 6% and 28% in HA, CF and IFA tests. These patients had no detectable cysts. This indicates that in postoperative patients a negative CF test may be of a better prognostic value in indicating absence of cysts than the HA and IFA tests.

Complement Fixation Tests↗
PubMed1986

Capture-ELISA for serum IgM antibody to respiratory syncytial virus.

A four-component solid-phase capture enzyme immunoassay was set up to test for serum IgM antibody to respiratory syncytial (RS) virus and was compared with immunofluorescence assay (IFA). A total of 128 young children with acute respiratory infections were studied. Thirty-six were shown to be RS virus-positive by the detection of RS virus in nasopharyngeal secretions and 92 were RS virus-negative. A serum specimen was collected after admission to the hospital (days 0-4) and a further specimen was obtained during days 10-14. Out of 36 RS virus-positive patients, 28 (77.7%) were found to be positive for IgM by both capture-ELISA and IFA. Out of 92 RS virus-negative patients 5 (5.4%) were IgM-positive. Four false-positive results were obtained by IFA due to the presence of rheumatoid factor. The capture-ELISA was shown to be a reliable technique in detecting specific IgM antibody to RS virus.

Antibodies, Viral↗
PubMed1975

The specific immunoglobulin response in cattle to Theileria parva (Muguga) infection.

Cattle were inoculated with dilutions of Theileria parva (Muguga) stabilate without concurrent antibiotic treatment. The dilutions were selected to induce severe East Coast fever infections, which in some cases would lead to death. Serum samples were collected and the complement fixation (CF), indirect haemagglutination (IHA) and indirect immunofluorescence (IFA) tests were used to detect T. parva serum antibodies. Sera were fractionated on Sephadex G200 to determine the activities of specific immunoglobulin fractions. Sequential production of IgM antibody followed by 7SIg antibody was demonstrated by the IHA and CF tests. All IFA activity was restricted to the 7SIg fraction. Serologic reactions in the IHA test were detected as early as 4 days after inoculation of the stabilate, CF activity after 8 days and IFA activity after 21 days.

Animals↗
PubMed1991

Sudden unexplained death syndrome--a new manifestation in melioidosis?

The indirect haemagglutination (IHA) test using sensitized turkey erythrocytes and the indirect immunofluorescence assay (IgM-IFA) was confirmed to be sensitive in the detection of a recent or current Pseudomonas pseudomallei infection in 19 culture-confirmed Singapore melioidosis patients. All were found to have antibody titres from 4 to 32768 in the IHA test and 10 to 320 in the IgM-IFA test. When these tests were employed on sera from 16 immigrant Thai construction workers who died of sudden unexplained death syndrome (SUDS) and 73 healthy Thai fellow workers, 93.8% and 68.8% of SUDS cases had IHA titre of greater than or equal to 4 and IgM-IFA titre of greater than or equal to 10 respectively, in contrast to 39.7% and 12.3% found among healthy Thai workers. These data indicate that at the time of death, most of the SUDS patients had an active infection with P. pseudomallei, possibly resulting from reactivation of a latent infection. The aetiological role of P. pseudomallei as the major cause of SUDS is discussed.

Adult↗
PubMed1995

Serosurvey and laboratory diagnosis of imported sandfly fever virus, serotype Toscana, infection in Germany.

Of eight acute infections in German tourists caused by sandfly fever virus, serotype Toscana (TOS), and diagnosed clinically and serologically, seven were acquired during visits to Tuscany, Italy, and one to Coimbra, Portugal. An indirect immunofluorescence assay (IFA) using infected cells, and a newly developed enzyme-immunoassay (EIA) using crude virus antigen prepared from infected Vero-E6 cells was used to detect anti-TOS IgM and IgG. In a seroepidemiological survey of 859 health care workers and medical students, anti-TOS IgG was detected in 1.0% by IFA, and in 0.7% by EIA. In 2034 German patients hospitalized for various diseases, 1.6% were positive for anti-TOS IgG by IFA, and 0.8% by EIA. Anti-TOS IgG was detected in 43 samples of commercial immunoglobulins at titres of 10-1000 by EIA. Although the seroprevalence of antibodies to TOS is low in Germany, TOS infection should be considered in patients returning from endemic areas who complain of fever, and headaches, and have symptoms of meningitis.

Adult↗
PubMed1996

Legionellosis linked with a hotel car park--how many were infected?

An outbreak of legionellosis associated with a hotel in Sydney, Australia, and the subsequent epidemiological and environmental investigations are described. Four cases of Legionnaires' disease were notified to the Public Health Unit. A cross-sectional study of 184 people who attended a seminar at the hotel was carried out. Serological and questionnaire data were obtained for 152 (83%) of these. Twenty-eight (18%) respondents reported symptoms compatible with legionellosis. Thirty-three subjects (22%) had indirect fluorescent antibody (IFA) titres to Legionella pneumophila serogroup 1 (Lp-1) of 128 or higher. The only site which those with symptoms of legionellosis and IFA titre > or = 128 were more likely to have visited than controls was the hotel car park (adjusted odds ratio [OR] 14.7, 95% confidence interval [CI]: 1.8-123.1). Those with symptoms compatible with legionellosis, but whose IFA titres were < 128 were also more likely to have visited the hotel car park (adjusted OR 4.4, 95% CI: 1.5-12.9). Seroprevalence of Lp-1 antibodies was higher in those who attended the seminar than in a population sample of similar age. Findings suggested that the 4 cases represented a small fraction of all those infected, and highlighted difficulties in defining illness caused by Lp-1 and in interpreting serology.

Adult↗
PubMed1998

Iris arteriovenous communication: clinical and angiographic features.

BACKGROUND: Iris arteriovenous communication (IAVC) represents a quite rare congenital anomaly, consisting of abnormal vascular connection bypassing the iris capillary bed. The aim of the present study is to describe clinical and angiographic pattern of IAVC on iris fluorescein angiography (IFA) and on indocyanine green videoangiography (IICGV). METHODS: During a mean follow-up period of 33.5 months, eight patients affected by IAVC underwent at least three ophthalmological examinations completed by IFA and IICGV. RESULTS: IFA allows the detection of IAVC vascular structures, evidencing afferent and efferent branches, which show a rapid filling, without any evidence of leakage or iris hypoperfusion. IICGV shows more precisely the entire vascular pattern of IAVC, revealing also the presence of iris hypoperfusion in the sector in which the IAVC lay. One patient underwent cataract surgery; three months later, two neovascular tufts appeared in the hypoperfused area related to IAVC. In all other patients, periodical examinations did not reveal any clinical or angiographic changes. CONCLUSION: In IAVC, the clinical picture appears stable throughout the follow-up; both angiographic techniques seem able to precisely delineate the vascular pattern. Nevertheless, IICGV is superior in showing iris hypoperfusion surrounding the vascular abnormality. Particular care must be drawn to patients affected by IAVC who need cataract surgery.

Arteriovenous Malformations↗
PubMed2002

Low prevalence of Bartonella henselae infections in Norwegian domestic and feral cats.

Bartonella henselae is the causative agent of cat scratch disease (CSD). This clinical entity is very rarely encountered in human medical practice in Norway. B. henselae infections including bacteraemia in cats have been frequently reported. The objective of the present study was to investigate the seroprevalence rate and the degree of B. henselae bacteraemia in Norwegian domestic and feral cats. One hundred cats investigated at a small animal veterinary practice in the middle of Norway were included in the study. Blood collected in Isolator blood-lysis tubes and lysates of erythrocytes after freezing and thawing were cultured. PCR analysis of whole blood was also performed. Serology was performed by indirect fluorescence assay (IFA) and enzyme immunoassay (EIA) using immobilised B. henselae Houston-1 strain as antigen. None of the 100 cats investigated was found to be bacteraemic. All 100 cats were seronegative when analysed by IFA; one cat was positive by EIA. The discrepancy between IFA and EIA of this particular cat is probably due to cross-reactive antibodies. Contrary to findings reported from several geographic regions, B. henselae infections in Norwegian cats appear to be virtually absent. This in turn may explain why CSD has not been reported in human medical practice in Norway.

Animals↗
PubMed2001

Immunization to Porphyromonas gingivalis enhances the local pro-inflammatory response to subcutaneous bacterial challenge.

BACKGROUND, AIMS: Human and animal studies have suggested that immunization to P. gingivalis might be beneficial for controlling periodontitis, by the induction of protective antibody response. The present study was designed to examine the effect of immunization on the local cellular, cytokine and antibody response to P. gingivalis in mice. METHODS: Subcutaneous chambers were implanted in 3 groups of mice. 2 groups were then immunized with P. gingivalis in either incomplete Freund's (IFA) or an Alum-based adjuvant. The 3rd group served as the control. At baseline, all mice were challenged with an intra-chamber injection of P. gingivalis. Chamber exudates were sampled at baseline, 1 and 7 days post-challenge, following by determination of leukocyte counts and the cytokines TNF-alpha, IFNgamma (pro-inflammatory) and IL-10 (anti-inflammatory). IgG levels to P. gingivalis were analyzed in both the exudates and serum. RESULTS: Leukocyte accumulation increased in the chambers over the study period and was more marked in the immunized groups. P. gingivalis challenge induced the expression of the tested cytokines in all groups. Levels of IFNgamma showed a significantly greater increase in the immunized groups on day 1 post-challenge. By day 7, the levels in the controls had reached those of the immunized groups. IL-10 levels were significantly higher in the control group compared to the immunized groups on day 1 and by day 7 they were reduced significantly in all groups to barely detectable levels. While there were no significant differences in TNF-alpha levels between IFA and control groups, they were significantly higher in the Alum group on day 0 and 7. Both immunization protocols induced anti-P. gingivalis IgG. The Alum group achieved the highest antibody levels, which were due to the increased expression of IgG1, a marker of a Th2-response. CONCLUSIONS: The results suggest that immunization to P. gingivalis results in enhanced expression of pro-inflammatory, tissue-destructive cytokines in the inflammatory site. The nature of the adjuvant used for immunization allows manipulation of the T-cell response, and alum was more effective in reducing the inflammatory response than IFA.

Adjuvants, Immunologic↗
PubMed1997

The protective efficacy of a liposomal encapsulated 30 kDa secretory protein of Mycobacterium tuberculosis H37Ra against tuberculosis in mice.

The immunoprotective efficacy of the 30 kDa secretory protein isolated from mid-log phase culture filtrate of Mycobacterium tuberculosis H37Ra was investigated using liposomes as adjuvant. Immunization of animals with the 30 kDa protein entrapped in liposomes prepared by a freeze-thaw method and absorbed on alum induced both cellular (monitored by T cell proliferation assay and cytokine secretion, viz. IL-2, IFN-gamma) and humoral (evaluated by ELISA) responses which were comparable to those induced in the 30 kDa IFA-immunized group. The protection induced by liposome-encapsulated 30 kDa secretory protein was slightly lower than that induced in 30 kDa IFA-immunized animals on the basis of higher survival rates and decreased viable counts of M. tuberculosis H37Rv in various organs (spleen, lung and liver) after 30 days of infection compared to the controls. The results strongly indicate that liposomes are an ideal vaccine delivery system which can effectively replace IFA for the delivery of the immunoprotective 30 kDa secretory protein of M. tuberculosis H37Ra.

Adjuvants, Immunologic↗
PubMed1996

KSHV antibodies among Americans, Italians and Ugandans with and without Kaposi's sarcoma.

A major controversy regarding Kaposi's sarcoma-associated herpesvirus (KSHV or HHV8) is whether or not it is a ubiquitous infection of humans. Immunoassays based on KSHV- and Epstein-Barr virus (EBV)-coinfected cell lines show that most US AIDS-KS patients have specific antibodies to KSHV-related antigens. We have developed a sensitive indirect immunofluorescence assay (IFA) based on an EBV-negative, KSHV-infected cell line, BCP-1. When we used this IFA assay, KSHV-related antibodies were found in 71-88% of serum samples from US, Italian and Ugandan AIDS-KS patients, as well as all serum samples examined from HIV-seronegative KS patients. Although none of the US blood donors examined were KSHV seropositive by IFA, intermediate and high seroprevalence rates were found in Italian and Ugandan control populations. Antibody kinetics showed that more than half of the AIDS-KS patients who were examined IgG-seroconverted before KS development, and antibody levels did not decline after seroconversion. For these patients, seropositivity rates increased linearly with time, suggesting that the rate of infection was constant and that the risk of developing KS once infected with KSHV is not highly dependent on the duration of infection. These data strongly suggest that KSHV is not ubiquitous in most populations and that the virus may be under strict immunologic control in healthy KSHV-infected persons.

AIDS-Related Opportunistic Infections↗
PubMed2001

Occurrence and molecular genotyping of Cryptosporidium spp. in surface waters in Northern Ireland.

AIMS: To investigate the incidence and genotype of Cryptosporidium parvum oocysts in drinking water sources in Northern Ireland for the period 1996-1999, and to compare conventional and molecular methods of detection. METHODS AND RESULTS: Four hundred and seventy-four waters were investigated by conventional methods, namely immuno-fluorescent antibody detection (IFA; 380) and immuno-magnetic separation-IFA (IMS-IFA; 94), of which 14/474 (3%) were positive. Two hundred and fourteen samples (214/474) were also investigated by PCR techniques, targeting both the 18S rRNA and TRAP-C2 genes, of which 11/214 (5.1%) were positive. These 11 samples were classified as genotype II following sequence analysis of the TRAP-C2 amplicon. CONCLUSIONS: This study demonstrated the low incidence of oocysts of C. parvum in water sources in Northern Ireland. SIGNIFICANCE AND IMPACT OF THE STUDY: Such molecular-based techniques offer a number of advantages over conventional detection methodologies, namely greater sensitivity and specificity as well as the ability to provide accurate genotyping data rapidly, which may be valuable in directing operational management in potential outbreak situations.

Animals↗
PubMed2000

Human antibody responses to Wuchereria bancrofti infective larvae.

Human IgG antibody responses to Wuchereria bancrofti third stage infective larvae (L3) surface and somatic antigens were studied by indirect immunofluorescence (IFA) and immunoblot with endemic Egyptian sera (n = 115) with the aim of identifying targets of protective immunity. Human sera variably recognized 14 major bands in L3 by immunoblot. The statistical significance of group differences in antibody prevalence was assessed by the chi-squared test. Children and young adults (aged 10-20 years) tended to have antibodies to more L3 somatic antigens than older adults, with significant differences for bands at 66, 60 and 5 kDa. Infected subjects had more consistent antibody responses to antigens at 55, 50 and 6 kDa than endemic normal subjects with negative serum filarial antigen tests, who are presumed to be uninfected. A 5 kDa antigen was preferentially recognized by the latter group. Antibodies to L3 surface antigens were equally prevalent in uninfected children (75%) and adults (90%) but less prevalent in people with microfilaremia (38%) than in amicrofilaremic subjects with or without filarial antigenemia (81%) (P < 0.001). IFA-positive sera showed significantly enhanced recognition of antigens at 66, 40 and 14 kDa in immunoblots relative to IFA-negative sera. Additional studies are needed to further characterize antigens identified in this study and to establish whether they are indeed targets of protective immunity in humans.

Adolescent↗
PubMed2001

The utility of serology in diagnosing candidosis in non-neutropenic critically ill patients.

This study was carried out to evaluate the utility of serological tests in the diagnosis of candidal infections in non-neutropenic critically ill patients. A prospective study was carried out in a 10-bed general intensive care unit; all patients with at least one organic sample with Candida spp. were included. A therapeutic-diagnostic algorithm was designed, and patients were treated or not, according to a classification. Blood samples were taken, and serological tests included: antigenaemia detection using two different commercial latex kits (Cand-Tec and Pastorex) and antibody detection by indirect haemagglutination (IHA) and indirect germ tube immunofluorescence (IFA). A total of 56% of antibody tests (IHA 45% and IFA 64%) and 26% of antigen tests (Cand-Tec 36% and Pastorex 17%) were positive. The sensitivity and specificity of these tests with respect to systemic candidosis were 37% and 78%, respectively, for antibodies, and 0% and 90% for antigens. There was statistical significance for mortality and low levels of antibodies; Candida glabrata was detected by IFA and Candida tropicalis by Cand-Tec. Serological tests may help to define the prognosis of these patients and to support the detection of specific Candida species.

Antibodies, Fungal↗
PubMed2002

Transmission of Babesia microti in Minnesota through four blood donations from the same donor over a 6-month period.

BACKGROUND: Babesiosis is a tick-borne zoonosis caused by intraerythrocytic protozoa. More than 40 US cases of Babesia microti infection acquired by blood transfusion have been reported. This report describes the identification of a transfusion-associated case of babesiosis and the subsequent identification of the infected blood donor and three other infected recipients of cellular blood components from three other donations by this donor. STUDY DESIGN AND METHODS: Serum specimens from the donors of blood that had been made into cellular components received by the index recipient and from other recipients of such components from the implicated donor were tested by the indirect fluorescent antibody (IFA) assay for antibodies to B. microti. Whole blood from IFA-positive persons was tested by PCR for B. microti DNA. RESULTS: IFA testing of serum from 31 of 36 donors implicated a 45-year-old man (titer, 1 in 256), whose donation had been used for RBCs. He likely became infected when bitten by ticks while camping in Minnesota in June 1999 and had donated blood four times thereafter. As demonstrated by PCR, he remained parasitemic for at least 10 months. Of the five other surviving recipients of cellular blood components from the implicated donor, three recipients (one for each of the three other donations) had become infected through either RBC or platelet transfusions. CONCLUSIONS: Babesiosis should be included in the differential diagnosis of posttransfusion febrile illness, and effective means for preventing transmission by blood transfusion are needed.

Aged↗
PubMed2006

A Bayesian approach for the evaluation of six diagnostic assays and the estimation of Cryptosporidium prevalence in dairy calves.

The prevalence of Cryptosporidium in calves and the test properties of six diagnostic assays (microscopy (ME), an immunofluorescence assay (IFA), two ELISA and two PCR assays) were estimated using Bayesian analysis. In a first Bayesian approach, the test results of the four conventional techniques were used: ME, IFA and two ELISA. This four-test approach estimated that the calf prevalence was 17% (95% Probability Interval (PI): 0.1-0.28) and that the specificity estimates of the IFA and ELISA were high compared to ME. A six-test Bayesian model was developed using the test results of the 4 conventional assays and 2 PCR assays, resulting in a higher calf prevalence estimate (58% with a 95% PI: 0.5-0.66) and in a different test evaluation: the sensitivity estimates of the conventional techniques decreased in the six-test approach, due to the inclusion of two PCR assays with a higher sensitivity compared to the conventional techniques. The specificity estimates of these conventional assays were comparable in the four-test and six-test approach. These results both illustrate the potential and the pitfalls of a Bayesian analysis in estimating prevalence and test characteristics, since posterior estimates are variables depending both on the data at hand and prior information included in the analysis. The need for sensitive diagnostic assays in epidemiological studies is demonstrated, especially for the identification of subclinically infected animals since the PCR assays identify these animals with reduced oocyst excretion, which the conventional techniques fail to identify.

Animals↗
PubMed1984

Comparison of immunity to malaria in Sudan and Indonesia: crisis-form versus merozoite-invasion inhibition.

Immunity to falciparum malaria was compared in two populations from malarious areas of southern Sudan and Flores, Indonesia. In Sudan, splenomegaly in adults was rare and anti-plasmodium indirect fluorescent antibody (IFA) titers were low to moderate, 1:1,280 being the modal titer. Sudanese serum was profoundly inhibitory to cultured Plasmodium falciparum, reducing incorporation of radiolabeled hypoxanthine by 63-93% and severely retarding intraerythrocytic parasite development, resulting in moribund crisis-form parasites and virtually no healthy schizonts. In Flores, 64% of the serum donors had splenomegaly greater than or equal to Hackett spleen grade 4 or 5, and the modal IFA titer was 1:10,240. Sera from Indonesia did not retard intraerythrocytic parasite development, but inhibited merozoite erythrocyte invasion 22-87%. Anti-merozoite activity did not correlate with IFA titers. The differences in principal modes of anti-parasitic activity suggest that immunity to malaria in Sudan is based on cell-mediated immune mechanisms associated with crisis forms, merozoite neutralization being of secondary importance. In contrast, malaria immunity in Flores appears to be principally based on anti-merozoite antibody, which does not cause crisis forms and allows for development of reduced numbers of healthy schizonts. This less efficient mechanism may lead to a continuous low-grade parasitemia, which could explain the high specific malaria antibody titers and adult splenomegaly in Flores as compared to Sudan. This latter approach to immunity, being less efficient than the former, apparently results in chronic malaria infections with associated high Ig titers and splenomegaly.

Animals↗