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PubMed1982

Ribonucleic acid antigen from the Reiter treponeme used in an ELISA for antibodies in syphilis.

Purified RNA from Treponema phagedenis biotype Reiter was used as antigen in an enzyme-linked immunosorbent assay (ELISA) for IgG antibodies in syphilis. The RNA ELISA was compared with the fluorescent treponemal antibody absorption test (FTA-ABS), the Treponema pallidum immobilization test (TPI), and with two ELISAs using purified flagellum from the Reiter treponeme as antigen (flagellum ELISA), and sonified Reiter treponeme culture as antigen (sonicate ELISA). A total of 729 sera from patients with and without syphilis were studied. The RNA ELISA had a lower sensitivity (P less than 0.01) in primary syphilis than the flagellum ELISA, the sonicate ELISA, and the FTA-ABS. In treated syphilis the RNA ELISA was also less sensitive than the TPI (P less than 0.01).

Animals↗
PubMed2006

Evaluation of F-actin ELISA for the diagnosis of autoimmune hepatitis.

OBJECTIVE: Antibodies to F-actin have been proposed to increase specificity in the diagnosis of autoimmune hepatitis (AIH). We compared the diagnostic value of a new F-actin enzyme-linked immunosorbent assay (ELISA) with the current gold standard of detection of smooth muscle antibodies by indirect immunofluorescence (SMA-IFT). METHODS: Archived sera of 47 patients with SMA positive AIH were tested with the F-actin ELISA and SMA-IFT. Prospectively collected sera of 123 patients with various liver diseases, 35 of whom had AIH, were analyzed by both assays. Different cutoff limits were considered for the F-actin ELISA (increments between 20 and 60 ELISA units) and SMA-IFT (titers of 1:40, 1:80, and 1:160). RESULTS: The F-actin ELISA had a sensitivity of 100% to detect all of 47 SMA positive AIH sera and the value of the ELISA units correlated with that of SMA titers (p < 0.0001). In prospective analysis, F-actin ELISA showed a superior sensitivity (74% vs 34%) and a similar specificity (98% vs 99%) and positive predictive value (88% vs 92%), compared with SMA-IFT. Combining both assays improved neither sensitivity nor specificity. CONCLUSIONS: The new F-actin ELISA seems to be a useful diagnostic tool with similar specificity and superior sensitivity for the diagnosis of AIH, compared with standard SMA-IFT. Due to its simplicity and operator independency, the F-actin ELISA may become a preferred screening technique for detection of autoantibodies in patients with suspected AIH.

Actins↗
PubMed1990

[Fundamental studies on serological diagnosis of amoebiasis. 1. Application of amoebic antigen for CF test to ELISA].

We studied the establishment of enzyme-linked immunosorbent assay (ELISA) using amoebic antigen for complement fixation (CF) test. Optimal dilution for ELISA of sera from patients was 1:100, and that of CF-antigen was 1:400. The upper limit of the 99% critical range of the reaction of negative sera was 0.068 (cut-off level). Absorbance of sera from patients diluted 1:100 to antigen and antibody titers of ELISA were strongly correlated, so it was possible to estimate antibody titers from absorbance of serum diluted 1:100. ELISA and CF test were done to compare sensitivity of the tests using 63 sera from patients with invasive amoebic disease. The sensitivity of ELISA compared well with CF test (62 sera were positive by ELISA and 61 by CF test). Only one sample was both positive by ELISA and negative by CF test. This sample had low ELISA titers, so this discrepancy was mainly due to the sensitivity of CF test in detecting lower levels of antibody. These results suggested that the amoebic antigen for CF test can be applicable to ELISA, and this method was so sensitive and specific.

Amebiasis↗
PubMed1991

Measurement of urinary lipopolysaccharide antibodies by ELISA as a screen for urinary tract infection.

Five hundred and twenty two clinical urine specimens submitted for routine microbiological examination were tested in parallel by conventional microscopy and culture and for lipopolysaccharide antibodies by an enzyme linked immunoabsorbent assay (ELISA) to assess the ELISA as a screen for urinary tract infection. When the ELISA alone was compared with routine methods the specificity sensitivity, and predictive value of positive and negative tests was 73.2%, 75.7%, 51.1% and 38.5%. For ELISA with microscopy the same variables were 71.1%, 82.2%, and 92.4% and 94.7%, respectively. The ELISA absorbency increased with increasing bacterial numbers, but results varied widely. Only 65.4% of urines which contained greater than or equal to 10(5) bacteria/ml were positive by ELISA; 36.8% of urines with less than 10(3) bacteria/ml were positive by ELISA; 100% of greater than or equal to 10(5) bacteria/ml cultures of Pseudomonas sp (n = 4), Staphylococcus aureus (n = 3), and Streptococcus faecalis (n = 2) were positive by ELISA but only 71.4% of Proteus sp (n = 7), 61.4% coliforms (n = 70), and 25% of coagulase negative staphylococci (n = 4). It is concluded that further development is required before the ELISA can be used for routine screening for urinary tract infection.

Antibodies↗
PubMed1988

Improved ELISA for thyroid microsomal auto-antibodies. Comparison with haemagglutination and immunofluorescent techniques.

Thyroid microsomal antibodies (TMA) and thyroglobulin antibodies (TGA) are strongly associated with auto-immune thyroid disease. TMA and TGA have been mostly detected by means of either immunofluorescence (IF), tanned red cell haemagglutination (TRCH), or radio-immunoassay (RIA) until the recent development of the enzyme-linked immunosorbent assay (ELISA). The ELISA has not been as extensively used in TMA detection as in the assay for TGA. The RIA method, though more sensitive, is technically and materially very demanding while the TRCH and IF are simpler to perform but are less sensitive. The main problem with the ELISA for the detection of TMA appears to be the interference of TGA present in some test sera reacting with thyroglobulin present as a contaminant in the thyroid microsomal preparation. In this study, we compared the TMA results from an ELISA system designed to eliminate TGA interference with those of IF and TRCH. The ELISA system in which TGA interference was eliminated without concurrent inhibition of the test reaction that gives rise to false negatives was more sensitive than either IF or TRCH. A significant number of samples was falsely reported as negative by both TRCH and IF. The correlation of the degree of positivity between TRCH and ELISA was moderate and was higher than that between ELISA and IF, though both were highly significant. The ELISA technique for TMA detection described here is a more efficient, more sensitive and also more cost-effective system than either TRCH or IF, both of which should now be replaced by ELISA, provided steps are taken to avoid false positives and false negatives.

Autoantibodies↗
PubMed2002

Field validation of a commercial blocking ELISA to differentiate antibody to transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus and to identify TGEV-infected swine herds.

A commercially available blocking ELISA was analyzed for its ability to identify antibodies to porcine coronaviruses (transmissible gastroenteritis virus [TGEV] or porcine respiratory coronavirus [PRCV]), to differentiate antibodies to TGEV and PRCV, and to identify TGEV-infected herds. Nine sera from uninfected pigs, 34 sera from 16 pigs experimentally infected with TGEV, and sera from 10 pigs experimentally infected with PRCV were evaluated using both the TGEV/PRCV blocking ELISA and a virus neutralization (VN) assay. The ELISA was not consistently effective in identifying pigs experimentally infected with TGEV until 21 days postinfection. Sera from 100 commercial swine herds (1,783 sera; median 15 per herd) were similarly evaluated using both tests. Thirty of these commercial herds had a clinical history of TGEV infection and a positive TGEV fluorescent antibody test recorded at necropsy within the last 35 months, while 70 herds had no history of clinical TGEV infection. The blocking ELISA and the VN showed good agreement (kappa 0.84) for the detection of porcine coronavirus antibody (TGEV or PRCV). The sensitivity (0.933) of the ELISA to identify TGEV-infected herds was good when considered on a herd basis. The ELISA was also highly specific (0.943) for the detection of TGEV-infected herds when the test results were evaluated on a herd basis. When sera from specific age groups were compared, the ELISA identified a greater proportion (0.83) of pigs in herds with TGEV antibody when suckling piglets were used. In repeatability experiments, the ELISA gave consistent results when the same sera were evaluated on different days (kappa 0.889) and when sera were evaluated before and after heating (kappa 0.888). The blocking ELISA was determined to be useful for herd monitoring programs and could be used alone without parallel use of the VN assay for the assessment of large swine populations for the detection of TGEV-infected herds.

Animals↗
PubMed1995

Evaluation and application of an indirect ELISA for the detection of antibodies to bovine respiratory syncytial virus in milk, bulk milk, and serum.

An indirect enzyme-linked immunosorbent assay (ELISA) was developed at the National Veterinary Institute (NVI), Uppsala, to detect antibodies to bovine respiratory syncytial virus (BRSV) in serum and milk. For the evaluation of the NVI ELISA, field sera collected from cattle in England and Sweden were tested in parallel with an ELISA in use at the Central Veterinary Laboratory (CVL), Weybridge. The tests showed 96% agreement. The sensitivity and specificity of the NVI ELISA relative to the CVL ELISA were 94% and 100%, respectively. There was evidence that the difference in sensitivity between the 2 tests was due to the detection of both IgG and IgM class antibodies by the CVL ELISA, whereas the NVI ELISA was designed specifically to detect IgG1. Milk and serum samples from individual cows were tested by the NVI ELISA for presence of antibodies to BRSV. There was a good correlation between the ability to detect antibodies in serum and the ability to detect them in milk, although the antibody titer was generally lower in milk than in serum. Bulk milk samples were collected from farms with severe clinical symptoms of respiratory distress and from farms with no history of respiratory disease. There was a clear distinction between antibody levels in diseased and healthy herds. The NVI ELISA is a rapid and reliable test for detecting antibodies to BRSV in milk, bulk milk, and serum samples.

Animals↗
PubMed1999

Evaluation of the baculovirus-expressed S glycoprotein of transmissible gastroenteritis virus (TGEV) as antigen in a competition ELISA to differentiate porcine respiratory coronavirus from TGEV antibodies in pigs.

The spike (S) glycoprotein of the Miller strain of transmissible gastroenteritis virus (TGEV) was recently cloned and expressed in baculovirus. The recombinant S protein was used as the coating antigen in a competition (blocking) enzyme-linked immunosorbent assay (ELISA) in combination with monoclonal antibodies to the S protein epitope A (conserved on TGEV and porcine respiratory coronavirus [PRCV]) or epitope D (present on TGEV only) to differentiate PRCV- from TGEV-induced antibodies. One set (set A) of 125 serum samples were collected at different times after inoculation of caesarean-derived, colostrum-deprived (n = 52) and conventional young pigs (n = 73) with 1 of the 2 porcine coronaviruses or uninoculated negative controls (TGEV/PRCV/negative = 75/30/20). A second set (set B) of 63 serum samples originated from adult sows inoculated with PRCV and the recombinant TGEV S protein or with mock-protein control and then exposed to virulent TGEV after challenge of their litters. Sera from set A were used to assess the accuracy indicators (sensitivity, specificity, accuracy) of the fixed-cell blocking ELISA, which uses swine testicular cells infected with the M6 strain of TGEV as the antigen source (ELISA 1) and the newly developed ELISA based on the recombinant S protein as antigen (ELISA 2). The sera from set B (adults) were tested for comparison. The plaque reduction virus neutralization test was used as a confirmatory test for the presence of antibodies to TGEV/PRCV in the test sera. The accuracy indicators for both ELISAs suggest that differential diagnosis can be of practical use at least 3 weeks after inoculation by testing the dual (acute/convalescent) samples from each individual in conjunction with another confirmatory (virus neutralization) antibody assay to provide valid and complete differentiation information. Moreover, whereas ELISA 1 had 10-20% false positive results to epitope D for PRCV-infected pigs (set A samples), no false-positive results to epitope D occurred using ELISA 2, indicating its greater specificity. The progression of seroresponses to the TGEV S protein epitopes A or D, as measured by the 2 ELISAs, was similar for both sets (A and B) of samples. Differentiation between TGEV and PRCV antibodies (based on seroresponses to epitope D) was consistently measured after the third week of inoculation.

Animals↗
PubMed2002

Characterization of inhibin forms and their measurement by an inhibin alpha-subunit ELISA in serum from postmenopausal women with ovarian cancer.

The aim of this study was to characterize the molecular wt forms of inhibins A and B and its free alpha-subunit present in serum from women with ovarian cancer as a basis for developing improved monoclonal antibody-based inhibin assays for monitoring ovarian cancer. Three new inhibin alpha-subunit (alphaC) ELISAs were developed using monoclonal antibodies directed to three nonoverlapping peptide regions of the alphaC region of the inhibin alpha-subunit. To characterize serum inhibin molecular wt forms present in women with ovarian cancer, existing inhibin immunoassays (inhibin A, inhibin B, and pro-alphaC) and the new alphaC ELISAs were applied to sera from women with granulosa cell tumors and mucinous carcinomas previously fractionated using a combined immunoaffinity chromatography, preparative SDS-PAGE, and electroelution procedure. The distribution and molecular size of dimeric inhibins and alpha-subunit detected were consistent with known mol wt forms of inhibins A and B and inhibin alpha-subunit and their precursor forms present in serum and follicular fluid from healthy women. The alphaC ELISAs recognized all known forms of inhibin and the free inhibin alpha-subunit, although differences between alphaC ELISAs were observed in their ability to detect high mol wt forms. To assess which of the alphaC ELISAs was preferred in application to ovarian cancer, the alphaC ELISAs were applied to serum from a range of normal postmenopausal women (n = 61) and postmenopausal women (n = 152) with ovarian (serous, mucinous, endometrioid, clear cell carcinomas, and granulosa cell tumors) and nonovarian (breast and colon) cancers. Despite differences in their ability to detect high mol wt forms of inhibin, the alphaC ELISAs showed similar sensitivity (i.e. proportion of cancer patients correctly detected) and specificity (proportion of controls correctly detected) indexes in the detection of mucinous carcinomas (84% and 95%) and granulosa cell tumors (100% and 95%) compared with earlier inhibin RIA or polyclonal antibody-based immunofluorometric assays. A combination of the alphaC ELISAs with the CA125 assay, an ovarian tumor marker that has a high sensitivity and specificity for other ovarian cancers (serous, clear cell, and endometrioid), resulted in an increase in sensitivity/specificity indexes (95% and 95%) for the all ovarian cancer group. These new monoclonal antibody-based inhibin alphaC ELISAs now provide practical and sensitive assays suitable for evaluation as diagnostic tests for monitoring ovarian cancers.

Adenocarcinoma, Mucinous↗
PubMed2002

Yolk protein immunoassays (YP-ELISA) to assess diet and reproductive quality of mass-reared Orius insidiosus (Heteroptera: Anthocoridae).

A yolk protein enzyme-linked immunosorbent assay (YP-ELISA) was developed for the predator Orius insidiosus (Say). The YP-ELISA is intended to assess reproductive response to dietary and other rearing conditions, and to assist in quality control and diet development for mass rearing. Hybridomas and monoclonal antibodies were produced against homogenates of eggs dissected from females. Hybridomas were selected for secretion of IgG that reacted with extracts of both females and their eggs, and that did not react with male extracts. Each cloned hybridoma produced a monoclonal antibody that specifically reacted on western blots against one of the two major yolk polypeptides, apoVn-I (180,000 molecular weight) or apoVn-II (40,000). Yolk protein ELISAs were developed with these antibodies to assess yolk protein content of female O. insidiosus as a measure of reproductive fitness and as a potential predictor of fecundity. Protocols for an indirect antigen ELISA and double antibody sandwich ELISA were developed to assess yolk protein contents of eggs and total contents in whole body homogenates. ELISA standards consisted of homogenates of eggs collected 0-24 h following oviposition. As determined with the sandwich ELISA, yolk protein contents of eggs declined with age before hatch, with a half-life of 32-34 h. Results were similar whether the detecting antibody-enzyme conjugate was anti-apoVn-I or anti-apoVn-II. Optimal conditions and sampling parameters were developed for the sandwich ELISA, which demonstrated minimal nonspecific interference in whole-insect extracts. In an initial application of the YP-ELISA, oviposition rates over a 10-d period were compared with yolk protein contents at the end of that period, dependent on diets of differing nutritional composition and quality. High and low yolk protein contents correlated with oviposition rates on respective diets, though oviposition showed more graded response to diets than did yolk protein. Improvements in sampling methods are discussed.

Animals↗
PubMed2006

Factors associated with ELISA scores for paratuberculosis in an Angus-Brahman multibreed herd of beef cattle.

Cow and calf genetic and environmental factors were evaluated for their association with ELISA scores for paratuberculosis in a multibreed population of beef cattle. The ELISA scores are a measure of the presence or absence of antibodies against Mycobacterium avium subsp. paratuberculosis in bovine serum. The linear mixed-model analysis used 352 ELISA scores from 238 cows: 51 Angus (A); 34 Brahman (B); 41 (3/4 A 1/4 B); 45 (1/2 A 1/2 B); 34 (1/4 A 3/4 B); and 33 Brangus (5/8 A 3/8 B). Cows were assumed to be unrelated. Year affected (P < 0.001) ELISA scores, but age of cow did not, which was expected to be significant because of the chronic progressive nature of this disease. Important regressions on fixed effects associated with cows were 1) a positive estimate of cow B breed effect (0.59 +/- 0.24; P < 0.017), indicating an upward trend of ELISA scores toward 100% B cows; 2) a negative estimate for weight change from before calving (late November) to the date of the blood sample in May (-0.0062 +/- 0.0019 score/kg; P < 0.002), indicating that poorer maintenance of cow weights was associated with higher ELISA scores; and 3) a positive estimate for days in lactation of cow on the date of the blood sample (0.0086 +/- 0.0034 score/d; P < 0.021), indicating the production of larger amounts of antibodies against Mycobacterium avium subsp. paratuberculosis as lactation progressed. Relevant regressions on fixed effects associated with calves were 1) calf birth weight (-0.022 +/- 0.010 score/kg; P < 0.035), and 2) calf gain from birth to the date of the cow blood sample (-0.0092 +/- 0.0027 score/kg; P < 0.001). These estimates indicate that cows that produced lighter calves at birth and/or calves with slower preweaning growth tended to have greater ELISA scores. Although the sensitivity (percentage of infected animals detected) of ELISA was only 50%, these results suggest that subclinical paratuberculosis may be negatively affecting cows and their offspring. Factors identified as associated with ELISA scores could help producers with culling decisions related to paratuberculosis control and eradication in beef cattle.

Animals↗
PubMed2004

[Inter-laboratory evaluation studies for development of notified ELISA methods for allergic substances (milk)].

Extracts of sausage, sauce, cookie, cereal and pasta sauce spiked with milk standard protein at a level of 5-20 ng/mL as sample solutions were analyzed in replicate in 10 laboratories. Coefficients of variation (CVs) of the three ELISA methods using a Milk Protein Casein ELISA Kit (Casein kit), a Milk Protein Beta-Lactoglobulin ELISA Kit (Beta-Lactoglobulin kit) and a FASTKIT Milk ELISA Kit (Milk ELISA kit) were mostly below 10%. Mean recoveries of the milk standard protein from the food extracts were over 40% in the three ELISA methods with a few excertions. The recoveries of milk standard protein from the sauce extract in Casein kit were improved by adjusting the extract to neutrality before the Casein kit assay. The recoveries of milk standard protein from cookie, cereal and pasta sauce were improved by the increasing the amount of antibody coated in the Milk ELISA kit. The detection limits of all the ELISA methods were 1 ng/mL in sample solutions. These results suggested that the notified ELISA methods are reliable and reproducible for the inspection of milk protein levels in extracts of sausage, sauce, cookie, cereal and pasta sauce.

Allergens↗
PubMed1985

Rapid serodiagnosis of leptospirosis using the IgM-specific Dot-ELISA: comparison with the microscopic agglutination test.

The dot enzyme-linked immunosorbent assay (Dot-ELISA) was compared to the microscopic agglutination test (MA test) for the diagnosis of human leptospirosis. Of 177 sera from 68 soldiers who trained in the Republic of Panama, 102 sera were positive in the MA test and 93 of these sera were positive in the IgM-specific Dot-ELISA. Incidence of infection was 50 of 68 patients with the MA test and 48 of 68 in the IgM Dot-ELISA. Five MA test-positive sera were reactive only in the IgG-specific Dot-ELISA, suggesting previous exposure. All 21 infecting serovars of Leptospira interrogans, as determined by positive reactions in the MA test or culture of blood and urine specimens, were reactive in the Dot-ELISA. Of 75 sera negative in the MA test, 61 were nonreactive in the Dot-ELISA. However, 9 of these 14 Dot-ELISA-positive/MA test-negative sera were acute samples from patients whose later sera were MA test-positive. Positive reactions in the IgM Dot-ELISA occurred in 2 of 30 control, 4 of 10 Lyme disease, 1 of 11 relapsing fever, and 1 of 8 yaws sera; 10 syphilis patient sera were nonreactive. The IgM-specific Dot-ELISA appears to be sensitive and specific for the serodiagnosis of acute leptospirosis. In addition, this rapid test is inexpensive, simple to perform, utilizes minute volumes of killed leptospiral antigen and is easily adaptable to field use.

Agglutination Tests↗
PubMed2000

Comparison of the sensitive screening kit with two ELISA sets for detection of anti-Toxocara antibodies.

Three serological tests (ELISA NOVUM, ELISA PU and ToxocaraCHEK) were compared using excretory-secretory (ES) antigen of second stage larvae of Toxocara canis on two sets of sera from patients aged 1-15 years, living in town and in the country, suspected of having larval toxocarosis. Of 60 serum samples examined for the presence of anti-Toxocara IgM and IgG antibodies 35 (58.3%) were positive with ELISA NOVUM, 30 (50.0%) with ELISA PU and 37 (61.6%) with ToxocaraCHEK. A significantly higher seropositivity with all the tests was detected in group of patients from the village (test chi2, p < 0.005) when compared with patients from the town. The highest sensitivity (100%) was detected for a rapid qualitative ToxocaraCHEK. Compared with this screening test, both ELISA sets showed higher specificity but a lower sensitivity. Consistent findings (+;-) with ELISA NOVUM and ELISA PU were 88.3%; ELISA NOVUM and ToxocaraCHEK 96.7% and ELISA PU and ToxocaraCHEK 86.7%. This comparison indicates the suitability of ToxocaraCHEK set for screening for its simplicity and rapidity in detecting anti-Toxocara antibodies.

Adolescent↗
PubMed1999

[Preliminary studies on microwave irradiation ELISA for diagnosis of Schistosomiasis japonica].

AIM: To explore a fast and highly efficient method for the diagnosis of schistosomiasis japonica. METHODS: Using microwave irradiation ELISA (MWI-ELISA) and fast-ELISA to detect specific antibodies in sera from 118 cases with schistosomiasis japonica, 61 healthy individuals and 12 paragonimiasis cases. RESULTS: The positive rates of schistosomiasis cases were 88.1% (104/118) by MWI-ELISA and 91.5% (108/118) by fast-ELISA, respectively (chi 2 = 0.74, P > 0.05). The false positive reaction of healthy individuals was found in 2 cases(3.3%) by MWI-ELISA and 1 case (1.6%) by fast-ELISA, respectively (chi 2 = 0.34, P > 0.05). No false positive reaction was found in paragonimiasis cases. CONCLUSION: The sensitivity and specificity of the two tests were similar, however, MWI-ELISA was faster than fast-ELISA.

Animals↗
PubMed2003

[Study on diagnosis of schistosomiasis by ELISA using periodate-treated soluble egg antigen].

OBJECTIVE: To improve SEA-ELISA, an immunodiagnostic assay for schistosomiasis. METHODS: Soluble egg antigen (SEA) of Schistosoma japonicum was treated with sodium periodate (SP) in order to oxidate its glycosylated epitopes. ELISA using the treated SEA was then performed to detect specific antibodies to SEA in the sera of schistosomiasis patients. RESULTS: Serum samples were tested by ELISA using SEA treated with sodium periodate (SP-SEA-ELISA), including 64 sera from cases with chronic schistosomiasis japonica, 119 sera from normal individuals in non-endemic area, 34 sera from patients with clonorchiasis, 33 sera of paragonimiasis cases and 36 sera from patients with cysticercosis. The results showed that its sensitivity (98.4%) was similar to that of the routine SEA-ELISA (100.0%) (P > 0.05) and the specificity is higher than that of the SEA-ELISA (P < 0.05). SP-SEA-ELISA showed a higher negative rate (89.0%) for sera of schistosomiasis patients 12 months post-treatment than that of the SEA-ELISA (42.1%). CONCLUSION: Use of SP-SEA can increase the specificity of ELISA, reduce cross-reactivity with serum samples from cases infected with other parasites and improve its value in evaluating therapeutic efficacy.

Animals↗
PubMed2004

Rapid ELISA for the diagnosis of rotavirus.

BACKGROUND & OBJECTIVES: Rotavirus is the major cause of gastroenteritis in infants and young children all over the world. The objective of the study was to develop a rapid ELISA for the diagnosis of rotavirus infection in children hospitalised with diarrhoea. METHODS: Immune serum was raised in rabbits by inoculating semipurified rotavirus, SA-11 strain. Immunoglobulins were conjugated to horse radish peroxidase and a rapid ELISA for rotavirus diagnosis was developed. The rapid ELISA was compared with routine ELISA, developed earlier at NIV. RESULTS: Of the 155 faecal samples from patients with diarrhoea, 96 were positive by rapid ELISA and 95 in routine NIV ELISA. OD values were higher in rapid ELISA. The rapid ELISA takes only 4 h to complete. INTERPRETATION & CONCLUSION: Rotavirus diagnosis by rapid ELISA is simple and easy to perform. This may lead to a significant reduction in the unnecessary usage of antibiotics, which cannot control infection due to rotavirus. This technology is being commercialized.

Child↗
PubMed1991

The dot-ELISA test for the detection of anti-PGL-1 IgM in leprosy patients and their contacts.

1. Since dot-ELISA has recently been reported to be a sensitive, simple and fast method, we have compared it with the conventional microplate ELISA method. Sera of 124 leprosy patients, 136 household and professional contacts, and 92 controls were tested for antibodies against a Mycobacterium leprae antigen using dot-ELISA on nitrocellulose membrane filters and microplate ELISA. 2. The sensitivity of the two techniques was similar for multibacillary patients, but dot-ELISA was less sensitive for paucibacillary patients although it was more specific (100%) than ELISA (93.4%). 3. Of 21 household contacts that gave a response by ELISA, 3 were also positive by dot-ELISA; one of these 3 developed indeterminate leprosy 12 months later and the other was diagnosed as borderline lepromatous after 28 months. 4. These data indicate that dot-ELISA has a high specificity and can be a useful tool in field evaluation.

Antigens, Bacterial↗