Dot Enzyme Immunoassay Research Articles

Negative ANA-IIF in SLE patients: what is beyond?

The antinuclear antibody (ANA) test has high sensitivity in diagnosing and classifying systemic lupus erythematosus (SLE).ObjectivesTo describe the immunological pattern of SLE patients through investigating specific antinuclear autoantibodies by enzyme dot immunoassay and studying their frequency in both positive and negative ANA indirect immunofluorescence assay (IIF) cases.MethodsIn a cross-sectional study, blood samples from 393 newly diagnosed SLE patients were analyzed using (IIF) on HEp-2 cells and ANA dot immunoassay by automated enzyme immunoassay (EIA) to detect 19 antibodies.ResultsNinety-one percent of the patients are females; their mean age was 37 ± 12.28. Antinuclear antibody (ANA) was detected by IIF in 82.4% of cases, with 181 (46.1%) speckled and 167 (42.4%) homogeneous ANA patterns. The majority of patients (96%) demonstrated autoantibodies via EIA. Among the ANA-IIF-negative patients, 97.2% demonstrated autoantibodies. There was a significant difference in the frequency of certain autoantibodies between SLE patients with negative and positive ANA-IIF (1.44 0.73, 3.12 2.09, p = 0.00) respectively.ConclusionThe results of analyzing 19 autoantibodies with the ANA staining pattern increased the significance of analyzing the immune profile even if IIF is negative when clinical symptoms strongly suggest SLE diagnosis. Certain autoantibodies may evade staining by the IFA approach while they are present in the patient’s serum, and they may not be detected by the ANA EIA profile if it does not contain that antigenic substrate.Key Points• Indirect immunofluorescence on Hep-2 is the conventional method for ANA detection and is regarded as the “gold standard” for testing in clinical practice for SLE.• In our study, ANA profile dot enzyme immunoassay (EIA)-based test was performed to evaluate 19 autoantibodies in SLE patients either positive or negative for ANA-IIF.• The presence of anti-dsDNA with ANA-IIF-negative serum in 32.4% of SLE patients provides evidence that not all anti-dsDNA antibodies are identified on standard HEp-2 substrates.• certain autoantibodies can evade staining by the ANA-IIF method despite being present in the SLE patient’s blood; this supports the ANA profile enzyme dot immunoassay as a more sensitive test.

First Report of Pea as a Natural Host of tomato mottle mosaic virus in China.

Tomato mottle mosaic virus, a new species in the genus Tobamovirus and family Virgaviridae, was first reported on tomato in Mexico in 2013 (Li et al. 2013). Subsequently, tomato mottle mosaic virus (ToMMV) was found infecting pepper and eggplant in China (Li et al. 2014; Chai et al. 2018). ToMMV was thought to pose a serious risk to solanaceous crops due to its potential to break resistance and numerous transmission ways (Li et al. 2020), meanwhile, some plant species in the families of Amaranthaceae, Brassicaceae, Cucurbitaceae, and Verbenaceae, were found as the hosts of ToMMV under experimental condition (Sui et al. 2017; Li et al. 2020). To clarify the occurrence of ToMMV in Yunnan province of China, 395 plant samples, including tomato (Solanum lycopersicum L.), pepper (Capsicum annuum L.) and pea (Pisum sativum L.), exhibiting viral-like symptoms were collected in major crop plantations in Yunnan province in 2020. Total nucleic acids were extracted from the diseased samples using a CTAB method (Li et al. 2008), and tested by reverse transcription polymerase chain reaction (RT-PCR) with ToMMV specific primers, ToMMVMPF (5'-ATGGCTCTAACTGTTAGTGGT-3') and ToMMVMPR (5'-TTAATACGAATCAGATCCCGCG-3'), which were designed based on the movement protein gene sequence of ToMMV YYMLJ isolate (KR824950). ToMMV was detected in 13 symptomatic samples (11 cherry tomato and 2 green pea plants) with a total detection rate of 3.29%. An 807-bp fragment was amplified from 2 out of 86 pea samples showing foliar chlorosis, mosaic, malformation and necrosis symptoms, and the amplicons were cloned and sequenced. Sequences obtained from the two pea samples were identical. Therefore, one sequence was deposited in the GenBank (accession no. MW561348). BLAST search result showed that the nucleotide sequence had the highest identity of 99.88% with the ToMMV TiLhaLJ isolate (KR824951). The ToMMV infection on the two pea samples was also verified by dot-enzyme immunoassay (Dot-ELISA) using ToMMV monoclonal antibody (kindly provided by Dr. Jianxiang Wu, Zhejiang University, China). To determine the pathogenicity of ToMMV on pea, ToMMV infectious cDNA clone was used to inoculate into 3-week-old healthy pea plants via an Agrobacterium-mediated method (Tu et al. 2021). Mottle or slight mosaic, chlorosis and malformation symptoms were observed on the upper leaves of the 8 out of 12 inoculated plants 8 days postinoculation, and ToMMV could be detected by RT-PCR from the 8 symptomatic plants but not from the asymptomatic and healthy control pea plants with the above described ToMMV specific primer pair. ToMMV has now only been detected in Mexico, China, Spain, USA, Israel and Australia on tomato, pepper and eggplant (Webster et al. 2014; Turina et al. 2016; Ambrós et al. 2017; Lovelock et al. 2020). To the best of our knowledge, this is the first report of natural infection of ToMMV in pea as well as the natural infection of ToMMV on plants outside of the family Solanaceae. The result also implies that ToMMV has a potential risk to more crops in the field. Since pea, pepper and tomato are economically important cash crops in China, proper virus management strategies for the cultivation of these crops should be adopted.

Eosinopenia in Patients With Typhoid Fever: A Case-Control Study

Background and objectivesSalmonella typhi is involved in one of the most prevalent infectious diseases of South East Asia, typhoid fever, but diagnostic tests cannot be performed regularly in developing countries. The objective of the study is to determine the association of eosinopenia with culture or serology-proven typhoid fever in patients, to determine the association of leukopenia with culture or serology-proven typhoid fever in patients and to determine independent predictors of eosinopenia.MethodsThis case-control study, done at Fauji Foundation Hospital, Pakistan, included patients with symptoms suggestive of typhoid fever, culture, or serology positive for typhoid fever and excluded patients who had been previously diagnosed with malaria and dengue hemorrhagic fever. After flagging cultures and serology, the records were viewed for leukocyte counts. Data, including age, gender, and clinical symptoms, were also recorded for analysis.ResultsOut of 200 participants, 59 participants with diagnosed typhoid fever had eosinopenia. There were 29 participants who had been diagnosed with typhoid fever via culture or serology and had leukopenia. Eosinopenia and leukopenia were more likely to be present in patients with a diagnosis of typhoid (OR: 9.60, 20.00). The independent predictors for eosinopenia were DOT enzyme immunoassay (DOT EIA) positive for Salmonella typhi, culture positive for Salmonella typhi and leukopenia (p<0.05).ConclusionThe presence of eosinopenia and features or serology suggestive of typhoid would most likely be associated with cultures being positive and, therefore, might provide an efficient means to begin treatment.

Retraction Note to: Antigen based detection of cystic echinococcosis in buffaloes using ELISA and Dot-EIA

[This retracts the article DOI: 10.1007/s12639-016-0762-9.].

An evaluation of a recombinant multiepitope based antigen for detection of Toxoplasma gondii specific antibodies

BackgroundThe inefficiency of the current tachyzoite antigen-based serological assays for the serodiagnosis of Toxoplasma gondii infection mandates the need for acquirement of reliable and standard diagnostic reagents. Recently, epitope-based antigens have emerged as an alternative diagnostic marker for the achievement of highly sensitive and specific capture antigens. In this study, the diagnostic utility of a recombinant multiepitope antigen (USM.TOXO1) for the serodiagnosis of human toxoplasmosis was evaluated.MethodsAn indirect enzyme-linked immunosorbent assay (ELISA) was developed to evaluate the usefulness of USM.TOXO1 antigen for the detection of IgG antibodies against Toxoplasma gondii in human sera. Whereas the reactivity of the developed antigen against IgM antibody was evaluated by western blot and Dot enzyme immunoassay (dot-EIA) analysis.ResultsThe diagnostic performance of the new antigens in IgG ELISA was achieved at the maximum values of 85.43% and 81.25% for diagnostic sensitivity and specificity respectively. The USM.TOXO1 was also proven to be reactive with anti- T. gondii IgM antibody.ConclusionsThis finding makes the USM.TOXO1 antigen an attractive candidate for improving the toxoplasmosis serodiagnosis and demonstrates that multiepitope antigens could be a potential and promising diagnostic marker for the development of high sensitive and accurate assays.

Age and Gender Difference of Typhoid Fever among Paediatric Patients Attended at a Tertiary Care Hospital in Bangladesh

Background: Typhoid fever varies according to different age and gender of the patients. Objective: The purpose of the present study was to observe different age and gender of typhoid fever patients among the paediatric group. Methodology: This cross sectional study was conducted at Department of Microbiology at Sir Salimullah Medical College, Dhaka, Bangladesh from January 2006 to December 2008 for a period of two (2) years. Blood samples from clinically suspected typhoid fever patients of different age and sex group was collected. Blood culture was done in all patients in the first week of illness. Widal test was done in all patients by using the first serum samples and in case of culture negative patients second serum samples (paired sera) were also tested by widal agglutination test. DOT Enzyme Immunoassay was also done in culture positive and culture negative typhoid patients by using the first serum sample. Result: A total number of 118(59%) paediatric patients were recruited. The highest rate of blood culture positivity for S. typhi was found among patients within 6 to 15 years of age group (19.14%). The rate of isolation of Salmonella typhi from blood of male patients were 19(15.1%) out of 126 cases and female patients were 11(14.9%) (P&gt;0.05). The highest number of patients was in the low income group (72.5%). Highest number, 37(63.8%) patients were within the 6-15 years of age group followed by 13(22.41%) in 16-25years age group. Conclusion: In conclusion typhoid fever mostly occurs in the age group of 6 to 15 years male patients with low socioeconomic status. Bangladesh Journal of Infectious Diseases 2016;3(2):36-39

Antigen based detection of cystic echinococcosis in buffaloes using ELISA and Dot-EIA.

Cystic echinococcosis is caused by the larval stage of the dog tapeworm, Echinococcus granulosus. The disease is recognized as one of the world's major zoonoses affecting human beings and domestic animals apart from its economic and public health importance. Development of the cysts in the intermediate host such as buffaloes occurs in the lungs, liver and other organs. In this study, detection of circulating antigen in the diagnosis of cystic echinococcosis in buffaloes was done using enzyme linked immunosorbent assay and Dot-Enzyme immunoassay (Dot-EIA). The sensitivity and specificity were determined as 89 and 92% respectively, whereas those of Dot-EIA were determined as 94 and 96%.

A Comparative Evaluation of Different Diagnostic Modalities in the Diagnosis of Typhoid Fever Using a Composite Reference Standard: A Tertiary Hospital Based Study in Central India.

Enteric fever, caused by Salmonella spp. is a major cause of morbidity and mortality worldwide and endemic in many developing countries including India and other South-East Asian countries. Blood culture is regarded as the gold standard for diagnosis. Currently, the standard serological method is tube agglutination with moderate sensitivity and specificity. Dot blot assay detecting IgM and IgG antibodies to a specific 50kD Outer Membrane Protein (OMP) antigen of Salmonella spp. is a simple, reliable, affordable and rapid test which can help in the early diagnosis of typhoid fever. To systematically evaluate the different diagnostic modalities against a composite reference standard for the better diagnosis of typhoid fever in clinically suspected cases of typhoid fever. This cross-sectional, prospective analytical study was carried out at a tertiary care hospital attached to Medical College in central India from November 2011 to June 2013. A total of 163 blood samples, collected aseptically from patients clinically diagnosed of enteric fever, were tested using various component tests like blood culture, Tube Widal and Dot Enzyme Immuno Assay (Dot EIA) for IgG and/or IgM. Composite Reference Standard (CRS) was created for defining the confirmed cases of typhoid fever using the component tests, wherein culture positive and in absence of culture positivity any two component test positive patients were taken as confirmed cases. All the component tests were evaluated against the CRS for sensitivity, specificity, PPV and NPV and their significance in relation to the duration of illness using statistical tests of significance. Blood culture was positive in 16 (9.81%) whereas, Tube Widal, IgM, IgG and IgM+IgG in combination were positive in 88(54%), 58(35.58%), 30 (18.40%) and 75 (46.01%) respectively. Using a two test criteria of CRS framed, a total of 104 patients were considered as confirmed cases. Though specificity of blood culture was 100%, the sensitivity was low with significant detection rate in 1st week of illness. Tube Widal showed a sensitivity of 65.38% and specificity of 89.83% with significant detection rate in 2nd week. Dot blot assay for IgM, IgG and Combined IgM and IgG showed a sensitivity of 71.15%, 65.28% and 51.72% respectively whereas, the specificity was 10.16%, 47.45% and 74.57% respectively with significant detection rate in 2nd and 3rd week of illness. It can be concluded that though blood culture is still the gold standard, Dot blot assay found to have high sensitivity and good specificity might be a practical alternative test for the rapid diagnosis of typhoid fever if interpreted with care particularly using a composite reference standard. Further, it is reliable, simple to perform and rapid; results being available in 1 hour when compared to 48 hours for blood culture and 18 hours for Tube Widal test.

Serodiagnosis of bovine fasciolosis by Dot-enzyme immuno assay in Chennai, Tamil Nadu

Excretory/secretory (E/S) antigen of adult Fasciola gigantica was used for diagnosis of fasciolosis in cattle and buffaloes sera from slaughter houses and different places in and around Chennai, Tamil Nadu. A total of 6.4mg of E/S antigen was obtained from the 60 flukes collected from slaughter houses. Coprological examination was done to detect endoparasitic infection in 321 fecal samples. Dot-enzyme immuno assay (Dot-EIA) was standardized with optimum working dilution of E/S antigen, serum, and conjugate by the checker board titration as 1 in 10 (50ng/ml), 1 in 20, and 1 in 1,000, respectively. A total of 321 sera samples consisting of 168 cattle and 153 buffaloes were tested by Dot-EIA. A total of 25 cattle (14.88%) and 38 buffaloes (24.83%) showed positive results in Dot-EIA. Sensitivity of Dot-EIA for cattle and buffaloes sera was 100%. Specificity of Dot-EIA for cattle and buffaloes was 95.28 and 92.10%, respectively in comparison to fecal examination by sedimentation method. Dot-EIA can be easily adapted in the field to detect fasciolosis in cattle and buffaloes.

Identification of Carriers Among Individuals Recruited in the Typhoid Registry in Malaysia Using Stool Culture, Polymerase Chain Reaction, and Dot Enzyme Immunoassay as Detection Tools

Chronic carriers of Salmonella Typhi act as reservoirs for the organism and become the agents of typhoid outbreaks in a community. In this study, chronic carriers in Kelantan, Malaysia were first identified using the culture and polymerase chain reaction method. Then, a novel serological tool, designated Typhidot-C, was evaluated in retrospect using the detected individuals as control positives. Chronic carriage positive by the culture and polymerase chain reaction method was recorded at 3.6% (4 out of 110) among individuals who previously had acute typhoid fever and a 9.4% (10 out of 106) carriage rate was observed among food handlers screened during outbreaks. The Typhidot-C assay was able to detect all these positive carriers showing its potential as a viable carrier screening tool and can be used for efficient detection of typhoid carriers in an endemic area. These findings were used to establish the first carrier registry for S Typhi carriers in Malaysia.

Leptospirosis Outbreak following Severe Flooding: A Rapid Assessment and Mass Prophylaxis Campaign; Guyana, January–February 2005

BackgroundLeptospirosis is a zoonosis usually transmitted through contact with water or soil contaminated with urine from infected animals. Severe flooding can put individuals at greater risk for contracting leptospirosis in endemic areas. Rapid testing for the disease and large-scale interventions are necessary to identify and control infection. We describe a leptospirosis outbreak following severe flooding and a mass chemoprophylaxis campaign in Guyana.Methodology/Principal FindingsFrom January–March 2005, we collected data on suspected leptospirosis hospitalizations and deaths. Laboratory testing included anti-leptospiral dot enzyme immunoassay (DST), immunohistochemistry (IHC) staining, and microscopic agglutination testing (MAT). DST testing was conducted for 105 (44%) of 236 patients; 52 (50%) tested positive. Four (57%) paired serum samples tested by MAT were confirmed leptospirosis. Of 34 total deaths attributed to leptospirosis, postmortem samples from 10 (83%) of 12 patients were positive by IHC. Of 201 patients interviewed, 89% reported direct contact with flood waters. A 3-week doxycycline chemoprophylaxis campaign reached over 280,000 people.ConclusionsA confirmed leptospirosis outbreak in Guyana occurred after severe flooding, resulting in a massive chemoprophylaxis campaign to try to limit morbidity and mortality.

Structural and functional studies of a 50 kDa antigenic protein from Salmonella enterica serovar Typhi

The high typhoid incidence rate in developing and under-developed countries emphasizes the need for a rapid, affordable and accessible diagnostic test for effective therapy and disease management. TYPHIDOT ®, a rapid dot enzyme immunoassay test for typhoid, was developed from the discovery of a ∼50 kDa protein specific for Salmonella enterica serovar Typhi. However, the structure of this antigen remains unknown till today. Studies on the structure of this antigen are important to elucidate its function, which will in turn increase the efficiency of the development and improvement of the typhoid detection test. This paper described the predictive structure and function of the antigenically specific protein. The homology modeling approach was employed to construct the three-dimensional structure of the antigen. The built structure possesses the features of TolC-like outer membrane protein. Molecular docking simulation was also performed to further probe the functionality of the antigen. Docking results showed that hexamminecobalt, Co(NH 3) 6 3+, as an inhibitor of TolC protein, formed favorable hydrogen bonds with D368 and D371 of the antigen. The single point (D368A, D371A) and double point (D368A and D371A) mutations of the antigen showed a decrease (single point mutation) and loss (double point mutations) of binding affinity towards hexamminecobalt. The architecture features of the built model and the docking simulation reinforced and supported that this antigen is indeed the variant of outer membrane protein, TolC. As channel proteins are important for the virulence and survival of bacteria, therefore this ∼50 kDa channel protein is a good specific target for typhoid detection test.

An improve dot enzyme immunoassay for serodiagnosis of melioidosis

An improve dot enzyme immunoassay for serodiagnosis of melioidosis

Comparative study of dot enzyme immunoassay (Typhidot-M) and Widal test in the diagnosis of typhoid fever

We compared the sensitivity and specificity of Typhidot-M and Widal test with blood culture (gold standard) for diagnosing typhoid fever in 105 children aged 1-15 years admitted with clinical suspicion of typhoid fever. Of the 105 cases, blood culture was positive for S.typhi in 41 (39%) children, Widal test was positive in 48 (45.7%) and Typhidot-M was positive in 78 (74.3%) cases. Sensitivity and specificity of Typhidot-M was 92.6% and 37.5% while sensitivity and specificity of Widal test was 34.1% and 42.8%, respectively. In children with fever of less than 7 days duration, Typhidot-M was positive in 97%, compared to 24.2% by Widal test. Typhidot-M is a simple and sensitive test for early diagnosis of typhoid fever in children.

Production and Characterization of Monoclonal Antibody Specific to Recombinant Dengue Multi-Epitope Protein

Monoclonal antibodies against novel dengue recombinant protein were produced following immunization of Balb/c mice with recombinant dengue multi-epitope protein (r-DMEP) expressed in Escherichia coli vector and purified in a single-step chromatography system. Antigenicity of r-DMEP was evaluated by dot enzyme immunoassay. Mice were immunized intraperitoneally with five doses each of 100 microg of this novel antigen at 1-week intervals and a final intravenous booster dose prior to the fusion. Hybridomas resulted from fusion of myeloma cells and splenocytes using PEG-1500 as an additive. Selection of the hybrids was done using HAT medium, and the hybrids thus selected were finally screened qualitatively and quantitatively by dot and plate immunoassays, respectively. Five antibody secretory hybrid clones exhibited specific reactivity against r-DMEP by dot-ELISA, whereas a lone clone was found to be cross-reactive with Japanese encephalitis virus (JEV). Monoclonal antibodies (MAbs) specific to r-DME protein recognized the envelope and non-structural epitopes by Western blot analysis. These MAbs were further checked for their diagnostic efficacy using dengue suspected clinical samples and found overall sensitivity and specificity for DRDE dipstick ELISA. MAb-based dipstick ELISA results were 85%, 75% and 85%, 90%, respectively.

Dot Enzyme Immunoassay (Typhidot(R)) in Diagnosis of Typhoid Fever in Children

Since typhoid fever continues to be a major health problem in developing countries emphasis is on for search of a simple rapid as well as sensitive and specific test/s which can be used in peripheral health services. Various serological test/s had been devised to detect antibodies and antigens of Salmonella enterica Serovar Typhi in the past. Ismail et al. detected a 50 kDa outer membrane protein (OMP) specific for the serotype Typhi and further evaluated the antigen using dot enzyme-immunoassay (EIA) format and observed that this antigen could react with Salmonella typhi-specific IgM and IgG in sera from patients with acute typhoid fever. Jesudason et al. reported Typhidot to be 100% sensitive and 80% specific with respect to blood culture a study from southern part of India. In the present study we performed dot EIA IgM and IgG (Typhidot) along with nested PCR Widal test and blood culture in eight pediatric patients (aged 1-12 years) which were suspected cases of acute typhoid fever in an endemic area. (excerpt)

Evaluation of tests for serological diagnosis of scrub typhus

Two specific serological tests, a Dot enzyme immunoassay (EIA) and an immunoglobulin (Ig)M enzyme-linked immunosorbent assay (ELISA) using the 56 kDa antigen and the Weil-Felix test were evaluated for diagnosis of scrub typhus. Sensitivity of 100, 86.5 and 43.5% were observed with Dot EIA, IgM ELISA and Weil-Felix test, respectively. False-positive reactions were observed in patients with falciparum malaria, pulmonary tuberculosis, S. viridans septicemia and typhoid fever using Dot EIA and IgM ELISA. Therefore, although Dot EIA and IgM ELISA are useful in the serodiagnosis of scrub typhus, efforts should be made to rule out other febrile illnesses.

A Dot-Enzyme Immunoassay for Large Scale Screening of Sera for Antibodies to Mycobacterium avium subsp. paratuberculosis in Goats

Rajukumar, K., Tripathi, B.N., Kurade, N.P. and Parihar, N.S. 2006. A dot-enzyme immunoassay for large scale screening of sera for antibodies to Mycobacterium avium subsp. paratuberculosis in goats. J. Appl. Anim. Res., 29: 49–52. A simple and effective dot-enzyme immunoassay procedure for large scale screening of antibodies to Mycobacterium avium subsp. paratuberculosis was evaluated using sequential sera from goats experimentally infected with M. a. paratuberculosis. Field evaluation of the test was carried, out on 168 serum samples from three goat flocks. The diagnostic value of the dot-EIA was comparable to that of plate ELISA. The sensitivities and specificities for the dot-EIA were found to be 65.6% and 92.9% and for plate ELISA 78% and 89%, respectively. Significant differences were not found in the positive and negative predictive values of both tests. The test design is suitable for screening of sera for antibodies to M. a. paratuberculosis at the field level.

Production of anti-dengue NS1 monoclonal antibodies by DNA immunization

Monoclonal antibodies against dengue NS1 protein were generated following immunization of mice with plasmid DNA encoding the transmembrane form of NS1 from dengue serotype 2 virus. A mammalian expression vector, pDisplay, was engineered to direct cell surface expression of dengue NS1 and tested for transient expression in COS cells. Two mice were immunized intramuscularly with six doses of 100 μg of plasmid at 2-week intervals; one mouse received a booster of live virus prior to the last plasmid injection. Both mice showed antibody responses against dengue antigens in dot enzyme immunoassay. Following fusion, hybridomas were screened with dot enzyme immunoassay against all four dengue serotypes. Specificity to the NS1 protein was confirmed by western blot analysis. Among five anti-dengue NS1 monoclonal antibodies generated, two clones were serotype 2 specific, two clones reacted with all four serotypes and the last also reacted with Japanese encephalitis virus. Reactivity against native or denatured forms of NS1 revealed three clones with reactivity to linear epitopes and two clones recognizing conformational epitopes. Such diverse specificity of anti-dengue NS1 monoclonal antibodies indicates that DNA immunization, especially with the combination of virus boosting, is an efficient way of producing monoclonal antibodies against viral protein. This has opened up a possibility of producing monoclonal antibodies to rare viral proteins that are difficult to isolate or purify.

Comparison of two immune diagnostic techniques for detection of Mycoplasma mycoides sub species mycoides SC in cattle

The use of Dot-Enzyme immunoassay (Dot-EIA) and Avidin-Biotin Complex (ABC-Elite) test in detecting Mycoplasma mycoides subspecies mycoides Small colony (MmmSc) in CBPP suspect tissues was evaluated. Out of the 330 suspect lung tissues and 20 suspect lymph nodes tissue samples analyzed, 245 (75.9%) and 276 (83.3%) of the lung tissues were positive by ABC-Elite and Dot-EIA respectively. However, the result of 11(3.45%) and 10(3.0%) of the lung tissues samples analyzed by ABC Elite and Dot EIA respectively could not be interpreted hence were considered to be doubtful. It is concluded that while both Dot-EIA and BC-Elite could be used efficiently in the field Dot-EIA is considered superior because of its cost-effectiveness, accuracy and speediness of results compared to ABC-Elite method. (Tropical Veterinarian: 2003 21(1): 23-29)