Comparison of an in-house latex agglutination test with IgM ELISA and MAT in the diagnosis of leptospirosis

The laboratory diagnosis of leptospirosis is fraught with several problems. Isolation of Leptospira by culture has a low sensitivity and the microscopic agglutination test (MAT) is time consuming To overcome these problems, a rapid latex agglutination test (LAT) has been standardized for the detection of antileptospiral antibodies in serum samples from suspected cases of leptospirosis. We compared the efficiency of the LAT to a commercially available IgM ELISA and MAT. A total of 150 serum samples were tested by LAT, IgM ELISA, and MAT. The positivity was 26.7%, 26% and 24% respectively. The sensitivity and specificity of LAT as compared to MAT was 90.62 and 91.96% respectively. Even though LAT and ELISA showed similar results, its rapidity and simplicity made latex agglutination test more suitable as a rapid screening test.     

A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos

ObjectivesTo compare two molecular assays (rrs quantitative PCR (qPCR) versus a combined 16SrRNA and LipL32 qPCR) on different sample types for diagnosing leptospirosis in febrile patients presenting to Mahosot Hospital, Vientiane, Laos.MethodsSerum, buffy coat and urine samples were collected on admission, and follow-up serum ∼10 days later. Leptospira spp. culture and microscopic agglutination tests (MAT) were performed as reference standards. Bayesian latent class modelling was performed to estimate sensitivity and specificity of each diagnostic test.ResultsIn all, 787 patients were included in the analysis: 4/787 (0.5%) were Leptospira culture positive, 30/787 (3.8%) were MAT positive, 76/787 (9.7%) were rrs qPCR positive and 20/787 (2.5%) were 16SrRNA/LipL32 qPCR positive for pathogenic Leptospira spp. in at least one sample. Estimated sensitivity and specificity (with 95% CI) of 16SrRNA/LipL32 qPCR on serum (53.9% (33.3%–81.8%); 99.6% (99.2%–100%)), buffy coat (58.8% (34.4%–90.9%); 99.9% (99.6%–100%)) and urine samples (45.0% (27.0%–66.7%); 99.6% (99.3%–100%)) were comparable with those of rrs qPCR, except specificity of 16SrRNA/LipL32 qPCR on urine samples was significantly higher (99.6% (99.3%–100%) vs. 92.5% (92.3%–92.8%), p <0.001). Sensitivities of MAT (16% (95% CI 6.3%–29.4%)) and culture (25% (95% CI 13.3%–44.4%)) were low. Mean positive Cq values showed that buffy coat samples were more frequently inhibitory to qPCR than either serum or urine (p <0.001).ConclusionsSerum and urine are better samples for qPCR than buffy coat, and 16SrRNA/LipL32 qPCR performs better than rrs qPCR on urine. Quantitative PCR on admission is a reliable rapid diagnostic tool, performing better than MAT or culture, with significant implications for clinical and epidemiological investigations of this global neglected disease.     

Detection of Antibodies to Babesia equi in Horses by a Latex Agglutination Test Using Recombinant EMA-1

A latex agglutination test (LAT) using recombinant equi merozoite antigen 1 (EMA-1) for the detection of antibodies to Babesia equi was developed. The LAT was able to differentiate very clearly between sera from B. equi-infected horses and sera from Babesia caballi-infected horses or from normal horses. The LAT results were identical to those of a previously developed enzyme-linked immunosorbent assay. These results indicate that LAT using recombinant EMA-1 might be very useful as a routine screening method for the diagnosis of B. equi infection.     

Development of an Enzyme-Linked Immunosorbent Assay Using a Recombinant LigA Fragment Comprising Repeat Domains 4 to 7.5 as an Antigen for Diagnosis of Equine Leptospirosis

Leptospira immunoglobulin (Ig)-like (Lig) proteins are a novel family of surface-associated proteins in which the N-terminal 630 amino acids are conserved. In this study, we truncated the LigA conserved region into 7 fragments comprising the 1st to 3rd (LigACon1-3), 4th to 7.5th (LigACon4-7.5), 4th (LigACon4), 4.5th to 5.5th (LigACon4.5–5.5), 5.5th to 6.5th (LigACon5.5–6.5), 4th to 5th (LigACon4-5), and 6th to 7.5th (LigACon6-7.5) repeat domains. All 7 recombinant Lig proteins were screened using a slot-shaped dot blot assay for the diagnosis of equine leptospirosis. Our results showed that LigACon4-7.5 is the best candidate diagnostic antigen in a slot-shaped dot blot assay. LigACon4-7.5 was further evaluated as an indirect enzyme-linked immunosorbent assay (ELISA) antigen for the detection of Leptospira antibodies in equine sera. This assay was evaluated with equine sera (n = 60) that were microscopic agglutination test (MAT) negative and sera (n = 220) that were MAT positive to the 5 serovars that most commonly cause equine leptospirosis. The indirect ELISA results showed that at a single serum dilution of 1:250, the sensitivity and specificity of ELISA were 80.0% and 87.2%, respectively, compared to those of MAT. In conclusion, an indirect ELISA was developed utilizing a recombinant LigA fragment comprising the 4th to 7.5th repeat domain (LigACon4-7.5) as a diagnostic antigen for equine leptospirosis. This ELISA was found to be sensitive and specific, and it yielded results that concurred with those of the standard MAT.     

Dépistage de la Leptospirose : performance de la trousse Sérion Elisa classic Leptospira IgM

Aim of the study

Evaluate the sensitivity and the specificity of anti-Leptospira IgM detection with the Serion Elisa classic kit in comparison to the Panbio Elisa kit.

Methods

Twenty-three sera of patients whom Taqman probe real time PCR is positive and 19 sera positive by the microscopic agglutination test (MAT) are studied for sensitivity. Specificity is evaluated with 49 pairs of sera negative by MAT and with sera positive in IgM to EBV, to syphilis, to Borrelia and positive to influenza virus antibodies.

Results

14/23 (61%) of the sera positive by PCR and 27/30 (90%) of the sera positive at significant level by MAT are positive by Serion IgM Elisa. Serogroups Icterohaemorrhagiae, Grippotyphosa, Australis, Tarassovi, Canicola, Sejroe, Patoc and Cynopteri are detected. 40/49 sera (82%) negative by MAT are negative by Serion IgM Elisa. Agreement with the Panbio kit is 61% (30/49). Results are discrepant in 19/49 cases corresponding to a positive result by Panbio IgM and to a negative result by Serion IgM. Serological cross-reactions are more frequent with IgM to syphilis and to Borrelia than with IgM to EBV and to influenza virus antibodies.

Conclusion

A similar sensitivity is observed between the two kits but specificity is higher with Serion. Positivity in IgM is 61% in cases of leptospirosis diagnosed by PCR.     

A latex agglutination test using a recombinant nucleoprotein for detection of antibodies against avian influenza virus

A latex agglutination test (LAT) was developed for detecting antibodies against avian influenza virus. The recombinant avian influenza virus nucleoprotein expressed in Escherichia coli was purified, coupled with latex beads, and used as an antigen for the LAT. The LAT was capable of detecting anti-avian influenza virus antibodies irrespective of the avian-influenza subtype, and in most cases, the results correlated with the results of an agar gel precipitation test (AGPT). However, in comparison with the AGPT, the LAT could detect the anti-avian influenza virus antibodies for a longer period of time after the infection. The nonspecific agglutination observed in uninfected chicken sera was resolved by pretreating the sera with dried chicken-liver powder for 1 h. The LAT is easy to perform, and even after considering the time required for pretreatment of the serum, the total time required for obtaining the results is reduced in comparison to the time required in the case of the AGPT. This easy and rapid LAT is considered to be useful for monitoring avian influenza virus infection in the field.     

Diagnostic accuracy of a rapid E1-antigen test for chikungunya virus infection in a reference setting

ObjectivesRapid diagnostic tests targeting virus-specific antigen could significantly enhance the diagnostic capacity for chikungunya virus infections. We evaluated the accuracy of an immunochromatographic antigen test for diagnosis of chikungunya in a reference laboratory for arboviruses.MethodsAn immunochromatographic rapid test that uses mouse monoclonal antibodies as a tracer against the E1-envelope protein of chikungunya (ARKRAY, Inc. Kyoto, Japan) was evaluated. Sensitivity was tested in sera from travellers with RT-PCR confirmed chikungunya virus infection (Eastern/Central/Southern African (ECSA) genotype) (n=9) and from patients diagnosed during the 2014-2015 chikungunya outbreak on Aruba (Asian genotype, n=30). Samples from patients with other febrile and non-febrile illnesses (n=26), sera spiked with Flavivirus and Alphavirus reference strains (n=13, including non-spiked serum), and samples containing other selected pathogens (n=20) were used to test specificity of the E1-antigen test.ResultsSensitivity of the E1-antigen test was 8/9 (88.9%, 95% CI 56.5–98.0) for the ECSA genotype, but only 10/30 (33.3%, 95% CI 19.2-51.2) for the Asian genotype. Overall diagnostic specificity was 49/59 (83.1%, 95% CI 71.5-90.5).ConclusionsThe E1-antigen test we evaluated had fair diagnostic sensitivity for ECSA genotype chikungunya, but low sensitivity for Asian genotype, and poor overall specificity. Antibodies that react across genotypes will be required for further development of a rapid test for chikungunya. Performance of new tests should be evaluated against different chikungunya genotypes.     

Pathogen-specific leptospiral proteins in urine of patients with febrile illness aids in differential diagnosis of leptospirosis from dengue

Leptospirosis and dengue are two commonly seen infectious diseases of the tropics. Differential diagnosis of leptospirosis from dengue fever is often difficult due to overlapping clinical symptoms and lack of economically viable and easy-to-perform laboratory tests. The gold standard for diagnosis is the microscopic agglutination test (MAT). In this study, the diagnostic potential of screening for pathogen-specific leptospiral antigens in urine samples is presented as a non-invasive method of disease diagnosis. In a study group of 40 patients, the serum was tested for anti-leptospiral antibodies by MAT and enzyme-linked immunosorbent assay (ELISA). Urine of these patients was screened for leptospiral antigens by ELISA using specific antibodies against LipL32, LipL41, Fla1, HbpA and sphingomyelinase. Group I patients (n = 23) were classified as leptospirosis-positive based on MAT and high titres of circulating IgM-specific anti-leptospiral antibodies. All of these patients excreted all five leptospiral antigens in the urine. The 17 MAT-negative cases included six patients with pyrexia of unknown origin (PUO; Group II) and 11 confirmed dengue patients (Group III). The latter tested negative for both serum anti-leptospiral antibodies and urinary leptospiral antigens. A salient outcome of this study was highlighting the usefulness of screening for urinary leptospiral antigens in disease diagnosis, as their presence confirmed leptospiral aetiology in two PUO patients. Immunoblots of urinary antigens identified well-defined bands corresponding to LipL32, HbpA and sphingomyelinase; the significance of the 42- and 58-kDa sphingomyelinase bands is discussed.     

Serodiagnosis of Equine Leptospirosis by Enzyme-Linked Immunosorbent Assay Using Four Recombinant Protein Markers

Leptospirosis, caused by Leptospira spp., is one of the most common zoonotic diseases in the world. We tested four recombinant proteins of Leptospira interrogans, namely, rLipL21, rLoa22, rLipL32, and rLigACon4-8, to evaluate their potential for use as antigens for the diagnosis of equine leptospirosis. We employed equine sera (n = 130) that were microscopic agglutination test (MAT) negative and sera (n = 176) that were MAT positive for the 5 serovars that most commonly cause equine leptospirosis. The sensitivity and specificity of ELISA compared to MAT were 82.39% and 86.15%, respectively, for LigACon4-8, 77.84% and 92.31%, respectively, for Loa22, 77.84% and 86.15%, respectively, for LipL32, and 84.66% and 83.85%, respectively, for LipL21. When one of the two antigens was test positive, the sensitivity and specificity of ELISA were 93.75% and 78.46%, respectively, for rLigACon4-8 and LipL32, 93.18% and 76.15%, respectively, for rLigACon4-8 and LipL21, 89.77% and 80.77%, respectively, for rLigACon4-8 and Loa22, 91.48% and 78.46%, respectively, for LipL21 and Loa22, 93.75% and 76.92%, respectively, for LipL21 and LipL32, and 90.34% and 80.77%, respectively, for Loa22 and LipL32. In conclusion, we have developed an indirect ELISA utilizing rLigACon4-8, rLoa22, rLipL32, and rLipL21 as diagnostic antigens for equine leptospirosis. The use of four antigens in the ELISA was found to be sensitive and specific, the assay was easy to perform, and the results concurred with the results of the standard Leptospira MAT.     

Diagnostic accuracy of tests for leprosy: a systematic review and meta-analysis

ObjectivesOwing to difficulties in the clinical diagnosis of leprosy, several complementary tests have been developed and used. The aim was to systematically summarize the accuracy of diagnostic tests for leprosy.MethodsWe searched for relevant articles in Embase, Medline, and Global Health databases, until June 2017. Studies evaluating the accuracy of any diagnostic techniques for differentiating between people with and without leprosy were included. Studies solely focusing on differentiating between the separate forms of leprosy were excluded. Our protocol was registered on PROSPERO (CRD42017071803). We assessed study quality using the QUADAS-2 checklist. A bivariate random effects regression model was used for the meta-analyses.ResultsWe included 78 studies, most of those evaluating the detection of IgM antibodies against phenolic glycolipid I using ELISA. Sensitivity of the 39 studies evaluating ELISA was 63.8% (95% CI 55.0–71.8); specificity 91.0% (95% CI 86.9–93.9). The lateral flow test (nine studies) and the agglutination test (five studies) had a slightly higher sensitivity and a slightly lower specificity. Sensitivity of qPCR was (five studies) 78.5% (95% CI 61.9–89.2) and specificity 89.3% (95% CI 61.4–97.8). Sensitivity of conventional PCR was (17 studies) 75.3% (95% CI 67.9–81.5) and specificity 94.5% (95% CI 91.4–96.5).ConclusionsAlthough the test accuracy looks reasonable, the studies suffered from heterogeneity and low methodological quality.     

The determination of measles antibody titers in the latex agglutination reaction]

A new latex agglutination test (LAT) for detection of antibodies to measles virus in blood sera was developed. The latex diagnosticum consists of polymeric microspheres of national make carrying on their surface covalently bound measles virus antigen. The data on the specificity and sensitivity of the test are presented, and the results of titrations of blood sera from schoolchildren by traditional HI test and LAT are compared. LAT was shown to be highly sensitive, much simpler than the available methods for detection of measles antibodies and more economic, and therefore may be recommended for mass surveys.     

Comparison between microscopic agglutination test (MAT) and enzyme-linked immunosorbent assay (ELISA) for detection of leptospiral antibodies in cattle

Leptospirosis is a worldwide zoonosis caused by Leptospira interrogans. It causes financial loss to the cattle industry with decreased milk production, abortion, stillbirth, infertility, and mortality. This study was conducted to evaluate two serological techniques, the microscopic agglutination test (MAT) and the enzyme-linked immunosorbent assay (ELISA), for detection of leptospiral antibodies in cattle. Three hundred and fifty five serum samples were collected from five commercial dairy farms in Tehran suburb from December 2004 to June 2005. The results of the MAT protocol showed that 55 serum samples (15.49%) had detectable antibodies against at least one serovar of L. interrogans at a dilution of ≥ 1:100. Icterohaemorrhagiae was the most (42.52%) and ballum the least (1.15%) prevalent serovars among 87 positive reactions against different serovars of L. interrogans. Leptospiral antibodies were also detected by the ELISA in 77 sera (21.69%) of 355 samples. The results of this study also showed that 2.25% of the sera that reacted positive in the MAT were negative by the ELISA, and 8.45% of the MAT negative sera were positive by the ELISA. Comparison between two techniques showed that there was a high correlation (0.61 < kappa?=?0.649 < 0.8) between the ELISA and MAT for detection of leptospiral antibodies in cattle.     

Bacteriophage-Based Latex Agglutination Test for Rapid Identification of Staphylococcus aureus

Rapid diagnosis is essential for the management of Staphylococcus aureus infections. A host recognition protein from S. aureus bacteriophage phiSLT was recombinantly produced and used to coat streptavidin latex beads to develop a latex agglutination test (LAT). The diagnostic accuracy of this bacteriophage-based test was compared with that of a conventional LAT, Pastorex Staph-Plus, by investigating a clinical collection of 86 S. aureus isolates and 128 coagulase-negative staphylococci (CoNS) from deep tissue infections. All of the clinical S. aureus isolates were correctly identified by the bacteriophage-based test. While most of the CoNS were correctly identified as non-S. aureus isolates, 7.9% of the CoNS caused agglutination. Thus, the sensitivity of the bacteriophage-based LAT for identification of S. aureus among clinical isolates was 100%, its specificity was 92.1%, its positive predictive value (PPV) was 89.6%, and its negative predictive value (NPV) was 100%. The sensitivity, specificity, PPV, and NPV of the Pastorex LAT for the identification of S. aureus were 100%, 99.2%, 98.9%, and 100%, respectively. Among the additionally tested 35 S. aureus and 91 non-S. aureus staphylococcal reference and type strains, 1 isolate was false negative by each system; 13 and 8 isolates were false positive by the bacteriophage-based and Pastorex LATs, respectively. The ability of the phiSLT protein to detect S. aureus was dependent on the presence of wall teichoic acid (WTA) and corresponded to the production of ribitol phosphate WTA, which is found in most S. aureus clones but only a small minority of CoNS. Bacteriophage-based LAT identification is a promising strategy for rapid pathogen identification. Finding more specific bacteriophage proteins would increase the specificity of this novel diagnostic approach.     

Use of Recombinant Flagellin Protein as a Tracer Antigen in a Fluorescence Polarization Assay for Diagnosis of Leptospirosis

The objective of the present study was to investigate the usefulness of a recombinant flagellar protein, FlaB, of Leptospira interrogans serovar pomona in the serodiagnosis of leptospirosis by the fluorescence polarization assay (FPA). The recombinant protein FlaB was purified to homogeneity by a combination of nickel-nitriloacetic acid agarose chromatography, electrophoresis, and electroelution. Purified FlaB was labeled with fluorescein isothiocyanate (FITC). Western blotting was performed by using bovine sera with microscopic agglutination test (MAT) titers of antibodies against L. interrogans serovar pomona and L. bergpetersenii serovars hardjo and sejroe to confirm the antigenicity of FlaB. Western blot analysis demonstrated that labeled as well as unlabeled FlaB was recognized by the positive sera tested, indicating the broad serovar cross-reactivity of this protein. It also indicated that labeling with FITC did not affect the antigenicity. By using FITC-labeled FlaB as a tracer antigen, a homogeneous FPA was developed to detect antileptospiral antibodies in bovine sera. A population of 208 MAT-positive and 208 MAT-negative serum samples was tested by FPA. The FPA cutoff was determined by receiver operating characteristic analysis. By FPA, 83.7% of the MAT-positive serum samples were positive and 81.2% of the MAT-negative serum samples were negative. Compared to the results of MAT, the positive predictive value of FPA was 81.7% and the negative predictive value of FPA was 83.3%. The FPA is a simple and rapid technique for the detection of anti-Leptospira antibodies.     

Detection of antibodies to Babesia equi in horses by a latex agglutination test using recombinant EMA-1

A latex agglutination test (LAT) using recombinant equi merozoite antigen 1 (EMA-1) for the detection of antibodies to Babesia equi was developed. The LAT was able to differentiate very clearly between sera from B. equi-infected horses and sera from Babesia caballi-infected horses or from normal horses. The LAT results were identical to those of a previously developed enzyme-linked immunosorbent assay. These results indicate that LAT using recombinant EMA-1 might be very useful as a routine screening method for the diagnosis of B. equi infection.     

Performance of a new rapid test for detecting anti-hepatitis E virus immunoglobulin M in immunocompetent and immunocompromised patients

BackgroundHepatitis E virus (HEV) infections are widespread in both developing and developed countries. It is, therefore, important to assess the performance of systems for rapidly detecting anti-HEV immunoglobulin M antibodies in human sera.ObjectivesTo evaluate the diagnostic value of a new immunochromatographic assay, the HEV IgM rapid test (Wantai).Study designBlood samples were taken from 30 acutely infected immunocompetent and 30 acutely infected immunocompromised patients, all with HEV RNA in their blood. Specificity and cross reactivity was assessed using samples from 30 HEV RNA negative immunocompetent patients who had acute Epstein–Barr virus (EBV) or cytomegalovirus (CMV) infections and 30 HEV RNA negative immunocompromised patients. The performance of the HEV IgM Rapid Test was compared to that of a conventional microplate enzyme immunoassay.ResultsThe sensitivity of the rapid test in immunocompetent patients was 90% (95% CI: 72.1–100%), similar to that of the Wantai microplate assay (sensitivity: 96.6%, 95% CI: 78.77–100%; p = 0.61). The sensitivity of the rapid test in immunocompromised patients was 73.3% (95% CI: 55.4–91.2%) and that of the microplate assay was 83.3% (95% CI: 65.44–100%; p = 0.53). The rapid test produced no false positive reactions with samples from HEV RNA negative patients; while the microplate assay gave two false positive results (3.3%).ConclusionThe new Wantai HEV IgM rapid test is easy to use and suitable for rapidly detecting acute hepatitis E infections in both immunocompetent and immunocompromised patients. However, HEV RNA must be detected using a molecular assay for diagnosing an HEV infection in anti-HEV IgM negative patients.     

Development and evaluation of a recombinant-glycoprotein-based latex agglutination test for rabies virus antibody assessment

The measurement of neutralizing antibodies induced by the glycoprotein of rabies virus is indispensable for assessing the level of neutralizing antibodies in animals or humans. A rapid fluorescent focus inhibition test (RFFIT) has been approved by WHO and is the most widely used method to measure the virus-neutralizing antibody content in serum, but a rapid test system would be of great value to screen large numbers of serum samples. To develop and evaluate a latex agglutination test (LAT) for measuring rabies virus antibodies, a recombinant glycoprotein was expressed in an insect cell system and purified, and the protein was coated onto latex beads at concentrations of 0.1, 0.25, 0.5, 0.75, and 1 mg/ml to find out the optimal concentration for coating latex beads. It was found that 0.5 mg/ml of recombinant protein was optimal for coating latex beads, and this concentration was used to sensitize the latex beads for screening of dog serum samples. Grading of LAT results was done with standard reference serum with known antibody titers. A total of 228 serum samples were tested, out of which 145 samples were positive by both RFFIT and LAT, and the specificity was found to be 100 %. In RFFIT, 151 samples were positive, the sensitivity was found to be 96.03 %, and the accuracy/concordance was found to be 97.39 %. A rapid field test—a latex agglutination test (LAT)—was developed and evaluated for rabies virus antibody assessment using recombinant glycoprotein of rabies virus expressed in an insect cell system.     

Cultural,histochemical, and immunohistochemical detection of pathogenic Leptospira species in the kidneys of cattle slaughtered in two abattoirs in Southwest Nigeria

ABSTRACT

The impact of leptospirosis on food animals and humans has not been fully elucidated. This study was designed to determine the prevalence of pathogenic Leptospira organism in renal tissues of slaughtered cattle from two major abattoirs in southwestern Nigeria. One hundred and seventy (170) kidney samples of different breeds, ages, and sexes were collected and examined using cultural isolation, microscopic agglutination test (MAT), Warthin silver (WSs) impregnation and immunohistochemistry (IH). Chi-square test was used with a confidence level set at 0.05 to ascertain associations between positive cases and sex, age, and breed. One hundred and thirty (76.5%) of the kidney samples were culturally positive while 83.3% (25/31), 41.6% (37/89), and 35% (7/20) were MAT, WSss, and immunohistochemically positive, respectively. Interstitial nephritis (87%, 94/108) and tubular nephrosis (77.8%, 84/108) were the most prominent and consistent microscopic lesions observed.

Serovars Hardjoprajitno (29%) and Bratislava (22.6%) were the most prevalent serovars. To the best of our knowledge, studies on determination of pathogenic Leptospira serovars using cultural isolation, MAT, WSSs, and IH from kidneys of naturally infected cattle in Nigeria are rare. This study is the most comprehensive work in the detection of pathogenic Leptospira species from cattle in Nigeria.     

Rapid Detection of Contagious Bovine Pleuropneumonia by a Mycoplasma mycoides subsp. mycoides SC Capsular Polysaccharide-Specific Antigen Detection Latex Agglutination Test

A latex agglutination test (LAT) has been developed for the diagnosis of contagious bovine pleuropneumonia (CBPP). The latex microspheres were coated with MmmSC polyclonal immunoglobulin G antiserum and detected MmmSC antigen in the serum of cattle infected with CBPP and in growth medium containing MmmSC. The specific antigen recognizsed by this test appeared to be the capsular polysaccharide (CPS). The LAT recognized all 23 strains of MmmSC examined in this study, with a sensitivity level of 2 ng of CPS, or the equivalent of 5 × 10³ CFU, in a reaction volume of 0.03 ml. Therefore, rapid identification of MmmSC cultures should be possible. Agglutination was also observed with the related goat pathogens and “Mycoplasma mycoides” cluster members Mycoplasma mycoides subsp. mycoides large colony biotype (four of six strains positive) and Mycoplasma mycoides subsp. capri (three of six strains positive), in agreement with the suggestion that these latter two mycoplasmas may in fact represent a single species (although collectively exhibiting two capsular serotypes). Comparisons in diagnosis with the complement fixation test (CFT) were made by using African field sera from CBPP-infected cattle. After 2 (or 3) min of incubation, the test detected 55% (or 61%) of CFT-positive sera and 29% (or 40%) of CFT-negative sera, with an overall correlation in diagnosis of 62% (or 61%). The rates for false-positive diagnoses made by using “known” CBPP-negative sera from the United Kingdom were 3 or 13% after 2 or 3 min of incubation, respectively. The data agree with previous findings that some CBPP CFT-negative misdiagnoses may occur due to “antibody eclipsing” by excess circulating antigen. The LAT combines low cost and high specificity with ease of application in the field, without the need for any specialist training or equipment.