RESUMO
Objective: In this study we evaluated the utility of Abortus Melitensis Ovis Suis Brucella PCR (AMOS PCR) for the molecular characterization of Brucella species and analyzed the associated risk factors for brucellosis in Central Indian and Meghalayan population. Methods: AMOS PCR was carried out in a total of 160 BSCP-31 PCR-positive DNA samples isolated previously from the blood of Central Indian (n = 90) and Meghalayan cohorts (n = 70). Clinical and associated risk factors recorded earlier were used to establish strain-specific disease outcomes in study cohorts. Results: Brucella melitensis was found to be the dominant strain in both Central Indian and Meghalayan cohorts (57.7% and 54.28%, respectively) followed by Brucella abortus (42.22% and 38.57%). Although rare, brucellosis cases in the Meghalayan population also showed the presence of Brucella suis (7.14%) and Brucella ovis (2.85%). Febrile illness was a major clinical risk factor in both study cohorts, while occupational risk factors like exposure to animals and raw milk consumption were major mediating factors for brucellosis in Central Indian cohorts. On the contrary, meat consumption was found to be significant predisposing factor for brucellosis in Meghalaya. Conclusion: Molecular characterization of Brucella species provides important public health data for mitigation, advocacy, and antimicrobial stewardship.
Assuntos
Brucelose , Brucelose/epidemiologia , Brucelose/microbiologia , Índia/epidemiologia , Humanos , Fatores de Risco , Masculino , Feminino , Adulto , Animais , Pessoa de Meia-Idade , Brucella/genética , Brucella/isolamento & purificação , Brucella/classificação , Brucella melitensis/genética , Brucella melitensis/isolamento & purificação , Reação em Cadeia da Polimerase , Adulto Jovem , AdolescenteRESUMO
PURPOSE: Human brucellosis is a neglected zoonotic disease of significant public health concern. Molecular diagnosis of brucella remains challenging in low resource settings, due to the high infrastructure and cost involved. Loop-mediated isothermal amplification (LAMP) is a rapid point of care polymerase chain reaction (PCR) with the utility of on-field molecular diagnosis and offers a convenient alternative to conventional PCR. In the present study, we developed and evaluated the diagnostic utility of in house LAMP PCR targeting the Brucella genus-specific bcsp-31 gene in patients having febrile illness. MATERIALS AND METHODS: The analytical sensitivity and specificity of bcsp-31 LAMP PCR was first evaluated using brucella (n â= â8) and non-brucella cultures (n â= â5), along with spiked clinical samples. The overall diagnostic utility of developed LAMP PCR was then further evaluated in 393 human samples suspected of brucellosis. RESULTS: The developed LAMP PCR could detect as low as 8 âfg of DNA by visual detection within 35min. We report sensitivity and specificity of the developed LAMP PCR as 90.91% and 99.37%.The accuracy of the developed test assay was found to be 98.60%. In clinical samples, LAMP gave positivity of 20% with the concordance of 89% with conventional PCR. CONCLUSION: To conclude, a rapid, efficacious, sensitive LAMP PCR targeting the bcsp 31 gene was developed. The existing LAMP PCR can be used as a point of care screening test in various low resource endemic setting in lieu of conventional PCR for estimation of prevalence data, diagnosis and treatment of brucellosis.
Assuntos
Brucella , Brucelose , Genes Bacterianos , Reação em Cadeia da Polimerase , Humanos , Brucella/classificação , Brucella/genética , Brucelose/diagnóstico , Brucelose/epidemiologia , Brucelose/microbiologia , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/normas , Sensibilidade e Especificidade , Testes Imediatos/normas , Técnicas de Diagnóstico Molecular/normas , Padrões de Referência , Fatores de Tempo , Prevalência , Zoonoses/diagnóstico , Zoonoses/epidemiologia , Zoonoses/microbiologia , Limite de DetecçãoRESUMO
Introduction: Brucellosis is a neglected zoonotic disease of major public health concern. In India, the incidence of brucellosis remains vastly underreported due to its non-specific clinical presentation and sub-optimal sensitivity of existing gold standard tests. Studies in Northeast India have shown high incidences of brucellosis in livestock, but the region lacks data on human brucellosis despite its high associated risk. In the present study, we report the seroprevalence of human brucellosis and its associated risk factors in Meghalaya, Northeast India. Materials and Methods: A prospective observational study was conducted in East Khasi Hills and Ri.Bhoi districts of Meghalaya, from July 2018 to July 2020. A total of 1046 suspected patients with febrile illness along with associated risk factors were recruited through camps and various diagnostic laboratories in the defined region as per the pre.specified inclusion and exclusion criteria. Baseline, demographics, and clinical characteristics were recorded of all the consenting participants. Blood samples were analyzed for brucellosis-specific IgM antibodies through enzyme-linked immunosorbent assay (ELISA). Results and discussion: The overall seroprevalence of brucellosis was found to be 11.37% in Meghalaya. Among recruited participants, females were found to be more susceptible than males. Risk factors such as consumption of meat were found to be more significantly associated with brucellosis disease in the study region. Among the clinical presentations, pyrexia of unknown origin, myalgia, and chronic fatigue syndrome were found to be significantly associated with brucellosis disease in IgM.positive cases. Conclusion: Our result suggests further epidemiological investigations for human brucellosis in Northeast India toward improved advocacy for accurate diagnosis, and development of proper response mechanism in areas of high endemicity.