ABSTRACT
The recombinant protein MSP5 has been established as an important antigen for serological diagnosis of Anaplasma marginale by enzyme-linked immunosorbent assay (ELISA). However, due to the high cost of specialized equipment, this technique is not accessible to all laboratories, especially in developing countries in areas where the disease is endemic. The present study describes the standardization of a latex agglutination test (LAT) to detect antibodies against A. marginale based on recombinant MSP5. Compared with indirect enzyme-linked immunosorbent assay (iELISA), the relative sensitivity and specificity of the LAT were 95.21% and 91.86% respectively, with an almost perfect agreement between tests (kappa index = 0.863). These results can be considered important for the serological diagnosis of A. marginale, as they indicate that the test represents a rapid and low cost alternative to ELISA.
Subject(s)
Animals , Cattle , Anaplasma marginale/immunology , Anaplasmosis/diagnosis , Antibodies, Bacterial/blood , Antigens, Bacterial , Bacterial Outer Membrane Proteins , Cattle Diseases/diagnosis , Diagnostic Tests, Routine/methods , Anaplasma marginale/genetics , Antigens, Bacterial/genetics , Bacterial Outer Membrane Proteins/genetics , Latex Fixation Tests/methods , Recombinant Proteins , Recombinant Proteins/genetics , Sensitivity and Specificity , Serologic Tests/methods , Veterinary Medicine/methodsABSTRACT
Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum' e 'Candidatus Mycoplasma turicensis' são os agentes causadores da micoplasmose felina, que podem causar anemia aguda ou crônica. O objetivo deste trabalho foi determinar a ocorrência de hemoplasmas em gatos domésticos de Belém, Pará. Para isso, 201 gatos foram divididos em três grupos: Grupo A foi composto por 101 gatos de rua capturados pelo Centro de Controle de Zoonoses, o grupo B foi composto por 62 gatos domiciliados e saudáveis e o grupo C foi composto por 38 gatos domiciliados que apresentavam alguma afecção clínica. Foram coletadas amostras de sangue para a realização de Reação em Cadeia pela Polimerase (PCR) para detectar o DNA destes agentes, os quais foram sequenciados e alinhados. A análise estatística foi realizada para detectar a associação entre a infecção, o sexo dos animais e os grupos experimentais. O DNA de pelo menos uma das espécies de hemoplasmas pesquisados foi detectado em 19,9% (40/201) das amostras, sendo o DNA de 'Candidatus M. haemominutum' encontrado em 7,96% (16/201) das amostras, M. haemofelis em 1,49% (3/201) das amostras, enquanto que o DNA de 'Candidatus M. turicensis' foi detectado em 12,93% (26/201) das amostras. O DNA destes três agentes foi detectado em gatos dos grupos A e C, enquanto que no grupo B foi detectado apenas 'Candidatus M. turicensis' e 'Candidatus M. haemominutum' Foi detectada a influência do sexo sobre a infecção hemoplasmas apenas entre 'Candidatus M. haemominutum' e machos. Estes resultados mostraram que os hemoplasmas circulam entre os gatos domésticos em Belém e 'Candidatus M. turicensis' e 'Candidatus M. haemominutum' foram mais comuns do que M. haemofelis, especialmente em gatos vadios.
Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum' and 'Candidatus Mycoplasma turicensis' are the causative agent of the feline mycoplasmosis, which could cause acute or chronic anemia. The aim of this work was to determine the occurrence of hemoplamas in domestic cats from Belém, Pará. To this, 201 cats were divided into three groups: Group A were composed by 101 stray cats captured by Zoonosis Control Center, group B were composed by 62 owners healthy cats and group C were composed by 38 owners cats that were suffering by some medical condition. Blood samples were collected to perform Polymerase Chain Reaction (PCR) to detect the DNA of these agents, which were sequenced and aligned. Statistical analysis was performed to detect association between the infection, the sex of the animals and experimental groups. The DNA of at least one of the hemoplasmas studied were detected in 19,9% (40/201) of the samples, being the DNA of 'Candidatus M. haemominutum' was found in 7.96% (16/201) of samples, M. haemofelis in 1.49% (3/201) of samples, while 'Candidatus M. turicensis' in 12.93% (26/201) of the samples. The DNA of these three agents was detected in cats from groups A and C, while in Group B was detected only 'Candidatus M. turicensis' and 'Candidatus M. haemominutum'. The influence of sex on hemoplasma infection was detected only between 'Candidatus M. haemominutum' and males. These findings showed that hemoplasma circulate among domestic cats in Belém, and 'Candidatus M. turicensis' and 'Candidatus M. haemominutum' were more common than M. haemofelis, especially in stray cats.