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PubMed1997

Study on the suitability of sow colostrum for the serological diagnosis of porcine reproductive and respiratory syndrome (PRRS).

Serum and colostrum from 73 sows were collected. The serum samples were tested by Immuno. Peroxidase Monolayer Assay (IPMA) and the corresponding colostrum samples with the indirect Immuno fluorescent Antibody (IFA) technique. All serum positive sows were colostrum positive and all colostrum negative were serum negative. Eight sows only reacted positively in the colostral testing. Compared to the serum standard test the specificity was 82.6% and the sensitivity 100%. The observed agreement between both tests was 89.2%. In addition all serum samples were also tested with the IF test (IFT). Of the eight sows which were negative in the IPMA serum test and positive in the IFA colostrum test, three were found positive when the serum was tested with IFA. Consequently, the observed agreement was higher at 93.2%. After the suitability of colostrum for porcine reproductive and respiratory syndrome (PRRS) diagnosis was demonstrated, 1915 colostrum samples collected from 135 different farms were tested in a comparative study with the IPMA and IFA techniques. Of the 1915 colostrum samples 139 were positive with both IPMA and IFA. With IPMA only, 43 samples were positive compared with 192 samples found positive with the IFA technique. A total of 1541 samples were negative in both tests. The observed agreement between both tests was 87.5%. The quotient of the observed agreement minus chance agreement and the maximum possible agreement beyond chance level (Kappa Quotient) was 0.49. In 90% of the farms that tested IFA positive there was a seroconversion of more than 50% of all colostrum tested. By comparison only 29% of the IPMA positive farms were positive with more than 50%. Based on the epidemiological findings on PRRS it was concluded that the IFA technique indicates a higher sensitivity for the detection of PRRS virus antibodies in sow colostrum. Finally the possible advantages and disadvantages of sow colostrum testing and serum testing are discussed.

Animals↗
PubMed1971

False-positive anti-Toxoplasma fluorescent-antibody tests in patients with antinuclear antibodies.

The indirect fluorescent-antibody (IFA) method for diagnosis of toxoplasmosis is widely used and is considered to be as specific as the Sabin-Feldman dye test. After observing a patient with systemic lupus erythematosus (SLE) who had a positive toxoplasma IFA test but a negative dye test, we studied sera with high titers of antinuclear antibodies from 16 SLE patients and from 2 with rheumatoid arthritis for Toxoplasma antibodies in the immunoglobulin G and M (IgG and IgM) IFA tests and the dye test. Results of these tests were compared with titers of antinuclear antibodies, precipitating antibodies to single-strand deoxyribonucleic acid (DNA), and binding antibodies by use of DNA labeled with (3)H-actinomycin D. Of 18 patients, 11 had IgG and 4 had IgM IFA Toxoplasma antibodies; only 2 had antibodies detectable in the dye test. The immunofluorescence patterns in the Toxoplasma IFA test were indistinguishable from those obtained in patients with toxoplasmosis without antinuclear antibodies. Absorption of SLE sera with DNA did not result in a decrease in Toxoplasma IFA titers. When SLE sera were absorbed with live T. gondii, a marked drop in IgG IFA titer was observed as well as a decrease in titers of antinuclear antibodies and (3)H-DNA binding. Treatment of Toxoplasma cells with deoxyribonuclease and ribonuclease did not decrease their fluorescence. These results suggest that T. gondii nuclear antigens can absorb antinuclear antibodies but do not have exposed substrates for deoxyribonuclease. Tests in which organisms containing "nuclear" antigens for IFA detection of antibodies to these organisms are used may result in "false-positives" with sera containing antinuclear antibodies.

Absorption↗
PubMed2001

Comparison of serologic assays for detection of antibodies against human herpesvirus 8.

Improvement of serologic assays for detection of antibodies against human herpesvirus 8 (HHV-8) is critical to better understand its epidemiology and biology. We produced the HHV-8 latent (ORF73) and lytic (ORF65, K8.1, and glycoprotein B) antigens in the Semliki Forest virus system and evaluated their performance in immunofluorescence assays (IFAs) and enzyme-linked immunosorbent assays (ELISAs). These assays were compared with other latent antigen-based assays, including an IFA based on primary effusion lymphoma (PEL) cells and an ELISA based on bacterially expressed ORF73 antigen, as well as with other lytic antigen-based assays, including an IFA based on induced PEL cells, a commercial ELISA based on purified virions, and ELISAs based on K8.1- and ORF65-derived oligopeptides. We used a panel of 180 serum specimens obtained from three groups expected to have high, intermediate, and low HHV-8 prevalences. Using three different evaluation methods, we found that (i) the performances of the lytic antigen-based ELISAs were almost equivalent, (ii) the lytic antigen-based assays were more sensitive than the latent antigen-based assays, and (iii) in general, IFAs were more sensitive than ELISAs based on the same open reading frame. We also found that serum specimens from healthy individuals contained antibodies cross-reactive with HHV-8 glycoprotein B that can potentially cause false-positive reactions in lytic PEL-based IFAs. Although this is not a substantial problem in most epidemiologic studies, it may confound the interpretation of data in studies that require high assay specificity. Because the K8.1-based IFA provides sensitivity similar to that of lytic PEL-based IFAs and improved specificity, it can be a useful alternative to the PEL-based IFAs.

Antibodies, Viral↗
PubMedDate not supplied

Comparison of indirect immunofluorescence test for measles antibodies with haemagglutination inhibition and plaque neutralization tests.

Indirect Immunofluorescence (IFA), Plaque Reduction Neutralization (PRN) and Haemagglutination Inhibition (HI) tests for measles antibodies were carried out in 197 sera obtained from umbilical cord and vaccinated children. The IFA was also applied to blood samples collected with filter paper. IFA results demonstrated that the test is relatively simple to perform, with good reproducibility for different antigen lots. Good correlation was obtained between IFA, PRN and HI antibody titers. Better correlation was demonstrated with IFA and PRN than with HI and PRN tests. Sensitivity of IFA in detecting antibody was less effective than PRN, however more effective than HI using rhesus monkey red blood cells. PRN antibody titers over 100 were detected by IFA but not by HI (9.7% with negative results). IFA may be of considerable practical use and able to substitute HI in seroepidemiological surveys and to evaluate vaccine efficacy. It also can be simplified by employing filter paper collected samples.

Adult↗
PubMed1999

Efficiency of indirect immunofluorescence assay as a confirmatory test for the diagnosis of human retrovirus infection (HIV-1 and HTLV-I/II) in different at risk populations

We compared the indirect immunofluorescence assay (IFA) with Western blot (Wb) as a confirmatory method to detect antibodies anti retrovirus (HIV-1 and HTLV-I/II). Positive and negative HIV-1 and HTLV-I/II serum samples from different risk populations were studied. Sensitivity, specificity, positive, negative predictive and kappa index values were assayed, to assess the IFA efficiency versus Wb. The following cell lines were used as a source of viral antigens: H9 ( HTLV-III b); MT-2 and MT-4 (persistently infected with HTLV-I) and MO-T (persistently infected with HTLV-II). Sensitivity and specificity rates for HIV-1 were 96.80% and 98.60% respectively, while predictive positive and negative values were 99.50% and 92.00% respectively. No differences were found in HIV IFA performance between the various populations studied. As for IFA HTLV system, the sensitivity and specificity values were 97.91% and 100% respectively with positive and negative predictive values of 100% and 97.92%. Moreover, the sensitivity of the IFA for HTLV-I/II proved to be higher when the samples were tested simultaneously against both antigens (HTLV-I-MT-2 and HTLV-II-MO-T). The overall IFA efficiency for HIV-1 and HTLV-I/II-MT-2 antibody detection probed to be very satisfactory with an excellent correlation with Wb (Kappa indexes 0. 93 and 0.98 respectively). These results confirmed that the IFA is a sensitive and specific alternative method for the confirmatory diagnosis of HIV-1 and HTLV-I/II infection in populations at different levels of risk to acquire the infection and suggest that IFA could be included in the serologic diagnostic algorithm.

Journal Article↗
PubMed2003

Polymerase chain reaction for the detection of Cryptosporidium spp. in cat feces.

The objective of this study was to compare a polymerase chain reaction (PCR) assay and a monoclonal antibody-based immunofluorescence assay (IFA) for detection of Cryptosporidium parvum in cat feces. Eight C. parvum-naive DSH cats were orally inoculated with 1 x 10(6) oocysts of a C. parvum human isolate. Fecal samples were collected before inoculation, daily for the next 30 days, and twice weekly until day 85. Methylprednisolone acetate was administered at 20 mg/kg i.m. on days 85, 92, and 99. From days 86 to 115, feces were collected daily and then up to twice weekly until day 126. Immunofluorescence assay was performed after collection of the samples, and then the samples were frozen at -70 C until assayed by PCR. Cryptosporidium parvum was detected by PCR in 101 of 353 samples and by IFA in 52 of 353 samples: 27 samples were PCR positive, IFA positive; 74 samples were PCR positive, IFA negative; 25 samples were PCR negative, IFA positive; and 227 samples were PCR negative, IFA negative. The percentage of concordance between IFA and PCR was 72%. Results of this study suggest that this PCR assay is more sensitive than IFA for detection of C. parvum in cat feces.

Animals↗
PubMedDate not supplied

Seroprevalence of Ehrlichia canis, Ehrlichia equi, and Ehrlichia risticii in sick dogs from North Carolina and Virginia.

Ehrlichia canis, E. equi, and E. risticii seroprevalence was determined by microimmunofluorescent antibody testing (IFA) in a sequential population of 1,845 sick dogs admitted during a 1-year period to the North Carolina State University Veterinary Teaching Hospital. A seroreactor was defined by a reciprocal IFA titer of > or =80 to E. canis, E. equi, or E. risticii antigens. Of the 48 IFA seroreactors, 44 dogs were seroreactive to E. canis, 21 to E. equi, and 0 to E. risticii. Seventeen dogs reacted to both E. canis and E. equi antigens. There was concordance of E. canis IFA and western immunoblot (WI) test results for 36/44 dogs. Because of cross-reactivity of E. canis sera with E. equi antigens, WI was of less utility to confirm E. equi exposure. After elimination of E. canis seroreactors, there was concordance of 2/4 E. equi IFA and WI test results. Based upon a retrospective review of medical records, ehrlichiosis was diagnosed in 10/48 (21%) IFA seroreactive dogs, 9 of which were confirmed positive by WI. Of the remaining 38 IFA seroreactors, 29 also were confirmed by E. canis or E. equi WI. These results indicate that (1) ehrlichiosis was not diagnosed in the majority of serologically confirmed cases, (2) based upon E. canis and E. equi WI analysis, IFA testing was not specific (21% false positive), (3) E. canis sera cross-react with E. equi antigens, and (4) serologic evidence of E. risticii infection was lacking in the dog population studied.

Animals↗
PubMed1996

Evaluation for detection of Cryptosporidium oocysts in diarrheal feces of calves.

For the detection of Cryptosporidium oocysts, fecal samples were collected from 201 calves which showed diarrhea. Among the 201 samples, 29 samples (14.4%) were positive for Cryptosporidium spp. by the DMSO-modified acid-fast stain (MAFS), 23 samples (11.4%) were positive by commercial kit (Meridian Diagnostics, Cincinnati, Ohio) and 23 by the indirect immunofluorescence antibody (IFA) assay employing the monoclonal antibody (mAb C6). When tested by both IFA and MAFS, 20 fecal samples were positive for Cryptosporidium oocysts whereas 169 fecal samples were negative. If the MAFS is considered a standard method for oocyst detection, the IFA showed 69% of sensitivity and 98% of specificity. When tested by both IFA and commercial kit, 22 fecal samples were positive for Cryptosporidium oocysts while 177 samples were negative. One sample tested by IFA was found to be false negative, when compared with the results by commercial kit. The sensitivity of IFA was calculated as high as 96%; the specificity as 99% and the predictive value was also 99%. In the present study, IFA employing the mAb C6 revealed that 23 samples (11.4%) were positive among the 201 calves showing diarrhea. Of 23 IFA positive samples, 4 samples (5%) showed cryptosporidial oocysts more than 10(5) OPG. Therefore, it is concluded that the calves showing cryptosporidial oocysts more than 10(5) OPG in the feces were highly associated with clinical cryptosporidiosis.

Animals↗
PubMedDate not supplied

Comparison of the immunofluorescent assay and reverse transcription-polymerase chain reaction to detect and type infectious bronchitis virus.

Indirect fluorescent antibody (IFA) assay and reverse transcription-polymerase chain reaction (RT-PCR) are two current methods commonly used for the detection of infectious bronchitis virus (IBV) and its serotypes. The objectives of this study were to compare the two methods relative to detecting IBV in chicken embryos that were artificially inoculated with Mass41, Ark99, or Mass41/Ark99 in serial embryo passages and in tracheas and cecal tonsils collected from vaccinated commercial flocks with and without respiratory problems. The tissues collected from these birds were used to inoculate chicken embryos. The chorioallantoic membranes collected from the inoculated embryos were examined by the IFA assay, and the allantoic fluids collected from the same embryos were subjected to the RT-PCR. The IFA assay and RT-PCR both were able to detect and type IBV when only one IBV strain was inoculated into embryos, regardless of the number of passages. When embryos were inoculated with Mass41 and Ark99 strains simultaneously, both strains were 100% detected by the RT-PCR. By the IFA assay, 20% of samples were positive for two strains and 80% were positive for the Ark only. Two of eight samples collected from healthy flocks that were vaccinated with Mass/Conn/Ark were positive by the IFA assay, and four of eight were positive by the RT-PCR. Sixteen percent (12/75) of the samples from birds experiencing respiratory problems were positive by the IFA assay and 35% (26/75) were positive by the RT-PCR. Results suggest that the RT-PCR is more sensitive than the IFA assay, especially when more than one strain of IBV is involved, but the IFA assay is rapid and cheaper than the RT-PCR.

Animals↗
PubMed2003

Detection of IgM specific antibody using indirect immunofluorescent assay for diagnosis of acute leptospirosis.

Leptospirosis is a worldwide zoonosis, caused by Leptospira interrogans. At the earlier stage of the disease, the IgM immunoassays are expected to have more sensitivity than other immunoassays. Previous reports showed that the indirect immunofluorescent assay for detection of immunoglobulins (IFA-Igs) against Leptospira spp showed higher sensitive and specific than some genus specific tests. The authors determined an efficacy of the indirect immunofluorescent assay for detection of IgM specific leptospiral antibody (IFA-IgM). One hundred and eighty patients with acute febrile illness without localizing signs admitted to Hat Yai Hospital were studied. Using the cut-off value of > or = 1:400, the overall sensitivity, and overall specificity of the IFA-IgM were 89.2 per cent and 95.1 per cent, respectively. They were slightly greater than those of the conventional IFA-Igs (86.5% and 91.6%). The first sera obtained from patients including acute sera and single sera showed a low sensitivity (32.4%) but still higher than the IFA-Igs (29.7%). None of the patients with various diseases commonly confused with leptospirosis or healthy blood donors gave a titer greater than 1:200. In conclusion, the IFA-IgM has the same protocol as the IFA-Igs but the sensitivity and specificity is slightly greater than the IFA-Igs. This is another alternative test for the diagnosis of acute leptospirosis.

Acute Disease↗
PubMed1991

Ten-year study comparing enzyme-linked immunosorbent assay with the immunofluorescent antibody test for detection of feline leukemia virus infection in cats.

Immunodetection tests for feline retroviruses are powerful tools used in modern veterinary practice. Veterinarians must fully understand the characteristics--strengths and weaknesses--of the FeLV tests so that the information gained from them can be used properly. Any FeLV ELISA or immunofluorescent antibody (IFA) test is a method for detection of FeLV infection (the virus) and is not a diagnostic test for leukemia or other feline disease. From previous studies, it was determined that the most accurate test for detection of persistent FeLV infection is the IFA test, which detects FeLV antigens in cytoplasm of leukocytes in the blood of infected cats. In the study reported here, 1,142,600 FeLV IFA tests were performed between June 1972 and December 1990. During this period 19.8% of the IFA test results were positive and 78% were negative. Evaluation was not possible for the remaining 2.2% of the tests because of lack of enough leukocytes in the smears to evaluate, or nonspecific staining reactions. In 1979, 7 years after introduction of the IFA test, in-hospital FeLV ELISA were introduced, which enabled veterinarians to test for FeLV in their hospitals. Ever since that time, continual discrepancies have been reported between results of FeLV ELISA and IFA tests, particularly between positive ELISA results and their IFA test confirmation. A 10-year comparison was made between practitioner-performed in-hospital FeLV ELISA (n = 20, 240 tests) results and FeLV IFA test performed by a commercial laboratory. All samples tested by ELISA were submitted (for confirmation of results) by veterinarians from the United States, Canada, Europe, Japan, and Australia.(ABSTRACT TRUNCATED AT 250 WORDS)

Algorithms↗
PubMed1990

Immunofluorescence, enzyme-linked immunosorbent assay, particle agglutination and western blot for the detection of antibody to human immunodeficiency virus type 1.

Immunofluorescence assay (IFA) has been applied for detection of antibody to human immunodeficiency virus type 1 (HIV-1). To compare the IFA with an enzyme-linked immunosorbent assay (ELISA) and particle agglutination (PA), we examined the antibody response to HIV-1 in 475 sera from AIDS, PGL and ARC patients as well as several risk groups and healthy persons by three methods. The positive results by any methods were confirmed by western blot (WB). The results by all methods were well correlated on the sera from 45 asymptomatic male homosexuals and 70 female prostitutes. There were some false positive results by ELISA in the sera from prisoners and healthy persons. Four sera from drug abusers were positive only by PA and IFA and were negative by ELISA. All were WB-inconclusive. Particle agglutination and IFA results were compared with western blot analysis on 208 ELISA-positive sera. All IFA-strongly positive sera (84%) were positive by western blot. The sera with weakly positive, negative and inconclusive results by IFA (16%) were possibly any of positive, inconclusive or negative by western blot. By PA, 200 of 208 (97%) sera were PA-positive and 1% of these sera were WB-inconclusive while the PA-negative sera were either negative or inconclusive by western blot. These results suggested that PA is a simple and sensitive method for screening of HIV-1 antibody while IFA could be a primary confirmatory test and western blot would then be used for confirming any IFA-negative or inconclusive results.

Agglutination Tests↗
PubMedDate not supplied

[Evaluation of indirect immunofluorescence as a supplementary test for the diagnosis of HIV-1 infection].

In order to be used as an alternative or complementary test to confirm HIV-1 infection, the efficiency of indirect immunofluorescence assay (IFA) was compared with Western blot (WB) in 362 samples from persons with high and low risk behaviour. A panel of sera with 220 WB positive, 122 WB negative and 20 WB indeterminate sera were tested by an "in house" IFA. The sensitivity of IFA was found to be 98.63% and the specificity 98.36%. Therefore, IFA appeared to be an efficient alternative method to WB, since the cost of testing by IFA is less than 10% of WB testing. We observed a direct relationship between WB protein reactivity and IFA results. In 15 samples with coincident indeterminate results for WB and IFA, antibody reactivity to p24 and gp160 presented the highest frequency. On the other hand, antibodies to viral glycoproteins were always present in IFA weak positive samples, showing their high predictive value.

AIDS Serodiagnosis↗
PubMed1999

Comparison of human sera reactivities in immunoblots with recombinant human herpesvirus (HHV)-8 proteins associated with the latent (ORF73) and lytic (ORFs 65, K8.1A, and K8.1B) replicative cycles and in immunofluorescence assays with HHV-8-infected BCBL-1 cells.

The development of reliable, sensitive, and specific serological methods for the detection of human herpesvirus-8 (HHV-8) antibodies is critical for a thorough understanding of HHV-8 prevalence and pathogenesis. To evaluate the potential usefulness of HHV-8 proteins in measuring the responses against both latent and lytic antigens, we selected 1 latent [open reading frame (ORF) 73] antigen and 3 HHV-8 lytic antigens (ORFs 65, K8.1A, and K8.1B) previously identified as immunogenic [Virology (1998) 243, 208-217]. Full-length genomic ORF 73 and full-length ORFs 65, K8.1A, and K8.1B from the cDNA clones were cloned, expressed in bacterial and baculovirus-insect cell expression systems, and purified as GST fusion proteins. These recombinant proteins were used in Western blot reactions to test sera from 104 human immunodeficiency virus (HIV)+/Kaposi's sarcoma (KS)+ homosexual men, 77 HIV+/KS- homosexual men, and 84 age-matched HIV-/KS- men. These sera were also tested in immunofluorescence assays (IFAs) with uninduced and 12-O-tetradecanoylphorbol-13-acetate-induced B cell lymphoma-1 cells to detect antibodies against latency-associated nuclear antigens (LANA) and antibodies against lytic antigens (cytoplasmic fluorescence). These sera exhibited differential reactivities reflecting different titers of antibodies against HHV-8 proteins, and variable reactivities were seen more commonly with the sera from HIV-/KS- adult men. In the Western blot assay, 89% (93 of 104) of HIV+/KS + sera, 60% (46 of 77) of HIV+/KS- sera, and 7% (6 of 84) HIV+/KS- sera were reactive with both latent and lytic recombinant antigens. Western blot reactions with ORF 73 protein were more sensitive than LANA-IFA results. The lytic IFA and lytic Western blot (ORFs 65 and K8.1A) assays were more sensitive than the ORF 73 Western blots and LANA-IFA. With an exception of 2 sera from the HIV-/KS- group, all sera positive for lytic IFA antibodies and ORF 65 and K8.1A antibodies were also positive for latent antibodies. With few exceptions, sera positive for ORF 65 antibodies were also positive for K8.1A antibodies, and sera recognized the K8.1A protein more often than the K8.1B protein. There is a high degree of concordance between IFA and Western blot reactions, suggesting that this panel of HHV-8 recombinant proteins could detect a majority of the HHV-8-seropositive individuals. These results suggest that IFA followed by confirmation with the Western blot reactions with a panel of latent and lytic immunogenic antigens would provide a reliable, sensitive, and specific method for the detection of HHV-8 antibodies.

Adolescent↗
PubMed1995

Presence of antibodies reacting with porcine circovirus in sera of humans, mice, and cattle.

Antibodies reacting with porcine circovirus (PCV) were found in sera of humans, mice, and cattle by means of an indirect immunofluorescence assay (IFA) and an ELISA. In man, the highest seroprevalence (23.9% in IFA and 30.2% in ELISA) was found among hospitalized patients with fever of partially unclear etiology. Non-hospitalized "healthy" persons of the former German Democratic Republic showed a significantly higher number of positive sera (IFA = 20%) than blood donors from Berlin-West (IFA = 8.6%). Murine sera reacted positive with PCV in IFA between 12 to 69% in different breeding groups and about 35% of cattle sera were found reactive with PCV in IFA. Double-staining IFAs, immuno-electron microscopy and immunoblotting showed that non-porcine antibodies reacted with PCV structural antigen. Mathematical analysis revealed that in ELISA, non-porcine antibodies reacted specifically with PCV. Loss of binding specificity of non-porcine antibodies in ELISA after storage of sera and lower maximal optical densities obtained at equal titers in ELISA with non-porcine than with porcine sera suggest that antibodies in man, mice and cattle are caused by related species specific viruses sharing antigenic epitopes with PCV.

Adult↗
PubMed1998

IgM to Bartonella henselae in cat-scratch disease and during acute Epstein-Barr virus infection.

The diagnostic value of IgM to Bartonella henselae was evaluated in 20 children with cat-scratch disease (CSD) and controls consisting of 20 blood donors and 20 children with enlarged lymph nodes without CSD by two indirect immunofluorescence assays (IFA). One was based on B. henselae cocultivated with Vero cells (host cell-associated IFA), and the other on B. henselae grown on agar (host cell-free IFA). With the host cell-associated IFA, 18 of 20 children with CSD revealed IgM, whereas only 14 did so with the host cell-free IFA. Sera of two blood donors as well as sera from three children with enlarged lymph nodes without CSD showed also positive IgM to cell-associated B. henselae. This study reveals that the IFA applied had sensitivities of 70-90% and specificities of 87.5-100% for detecting IgM to B. henselae. Additionally, 20 patients with IgM to Epstein-Barr virus (EBV) capsid antigen were tested for IgM to B. henselae. Sera of 16 and 9 of these patients revealed IgM to B. henselae with the host cell-associated and the host cell-free IFA, respectively. Using Western blot these sera were demonstrated to react against linearized proteins of Vero cells and of B. henselae. Thus, since acute EBV infection may substantially reduce the specificity of B. henselae-specific IgM tests, we conclude that diagnosis of CSD should be confirmed by a significant IgG titer to B. henselae or by detection of this pathogen.

Animals↗
PubMed1991

Diagnosis of measles by fluorescent antibody and culture of nasopharyngeal secretions.

An indirect fluorescent antibody test (IFA) was evaluated using commercial mouse anti-measles monoclonal antibody and FITC-labeled goat anti-mouse immunoglobulin. For measles isolation, specimens were inoculated into Rhesus monkey kidney (RMK) cells and, when available, CV-1 cells. 381 specimens were tested by IFA and 408 specimens were cultured from patients suspected of having measles. For the 381 specimens tested by both methods, IFA and culture were positive for 31%, culture alone for 14%, IFA alone for 15%, and both negative for 40%. This study indicates that both IFA and culture are required for maximum measles virus detection. Of the positive specimens, 48% were detected either by IFA only (24%) or culture only (24%). IFA was positive in 69% of the culture-positive specimens and therefore, provided rapid diagnosis for many patients.

Antibodies, Monoclonal↗
PubMed1991

Comparison of rapid immunofluorescence assay to cell culture isolation for the detection of influenza A and B viruses in nasopharyngeal secretions from infants and children.

In the hospital setting it is often critical to isolate patients appropriately in order to prevent nosocomial infection. This is especially true with respiratory infection in infants and young children. At the present time a rapid immunofluorescence assay (IFA) for respiratory syncytial and parainfluenza viruses is routinely carried out in our laboratory. During January and February of 1990 we used monoclonal antibodies specific for influenza A and B viruses (Baxter-Bartels, Bellevue, WA) in this rapid IFA. 152 samples of NPS were tested by cell culture isolation (CCI) and IFA for the presence of influenza antigens. Twenty-seven samples were positive by both methods, and 114 were negative by both. Three samples were positive by IFA and negative by CCI, while eight samples were positive by CCI and negative by IFA. Five of these eight samples were not positive until 10 to 14 days after inoculation into cell culture, suggesting that the virus inoculum was small. Using CCI as the 'gold' standard, IFA was 90% sensitive and 93% specific. Because of its turn-around time (2-4 h) and acceptable sensitivity and specificity, IFA for influenza viruses will be a routine test in our diagnostic laboratory during the influenza season.

Animals↗