Álvarez-Ojeda MG, Saldaña-Fuentes C, Ballesteros-Elizondo MR, Martínez-Vázquez IO, López-Merino A, Briones LE, Morales-Loredo A

Language: Spanish
References: 27
Page: 620-627
PDF size: 714.29 Kb.

ABSTRACT

Objective: The aim of this study was to evaluate the sensitivity and specificity of polymerase chain reaction for detection of Brucella spp in human blood samples compared with the serological tests and blood culture. Material and Methods: In 2005, a total of 92 people were sampled from the towns of Anahuac and Sabinas Hidalgo, Nuevo Leon, where an outbreak of human cases had taken place in the same year as this study. The sera collected were analyzed by serological tests according to the NOM 022-SS2-1994. DNA was obtained using CTAB extraction method and it was used to amplify a fragment of 223 bp of the coding sequence for a protein of 31 kDa present in all Brucella species. Results: The polymerase chain reaction test detected 23 positive samples. The sensitivity and specificity compared with RB was 44.68 and 95.56%, respectively. Compared with mouse antibody production, it was 51.61 and 88.52%, and 2-mercaptoethanol was 53.57 and 87.50%. When isolation (positives cultures) was compared with polymerase chain reaction, we obtained 100.0% sensitivity and 80.23% specificity, taking into account people with positive and negative serology. Conclusions: The polymerase chain reaction test can be an alternative tool to bacterial culture in human brucellosis diagnosis.

REFERENCES

1. Casañas MC, Queipo-Ortuño MI, Rodríguez-Torres A, et al. Specificity of a polymerase chain reaction assay of a target sequence on the 32-kilodalton Brucella antigen DNA used to diagnose human brucellosis. Eur J Clin Microbiol Infect Dis. 2001;20:127-31.

2. Ortiz EE, Silva E, Izquierdo M. Normalización y evaluación del inmunoensayo ABICAP-BRU para su diagnóstico serológico de la brucelosis bovina. Revista electrónica de veterinaria 2007;VIII:(4)57-89.

3. SINAVE/DGE/SALUD 2011 Boletín SSA. Vigilancia Epidemiológica Semana 39; 2011. [Consultado 4 abril 2012] Disponible en http://www. dgepi.salud.gob.mx/boletin/2011/sem39/pdf/cua8.pdf.

4. Briones-Lara E, Saucedo-Palacios G, Martínez-Vázquez IO, et al. Respuesta al tratamiento de brucelosis en niños. Evaluación con reacción de Huddleson y PCR. Rev Med Inst Mex Seguro Soc. 2007;45:448-55.

5. Al-Tawfiq JA. Therapeutic options for human brucellosis. Expert Rev Anti Infect Ther. 2008;6:109-20.

6. Pappas G, Akritidis N, Bosilkovski M, et al. Brucellosis. N Engl J Med. 2005;352:2325-36.

7. Secretaría de Salud. Norma Oficial Mexicana. NOM 022-SSA2-1994 para la prevención y control de la brucelosis en el hombre en el primer nivel de atención. Diario Oficial de la Federación; 30 noviembre 1995.

8. Vrioni G, Pappas G, Priavali E, et al. An Eternal Microbe: Brucella DNA Load Persists for Years after Clinical Cure. Clin Infect Dis. 2008;46:131-6.

9. Kazemi B, Yousefi Namin SA, Dowlatshahi M, et al. Detection of Brucella by Peripheral Blood PCR and Comparison with Culture and Serological Methods in Suspected Cases. Iranian J Publ Health. 2008;37: 96-102.

10. Marianelli C, Graziani C, Santangelo C, et al. Molecular Epidemiological and Antibiotic Susceptibility Characterization of Brucella Isolates from Humans in Sicily, Italy. J Clin Microbiol. 2007;45:2923-8.

11. Yu WL, Nielsen K. Review of detection of Brucella spp. by polymerase chain reaction. Croat Med J. 2010;51:306-13.

12. Milian-Suazo F. Manual para determinar el tamaño de muestra para estudios de campo en medicina veterinaria. México. INIFAP-Produce; 2001.

13. López-Merino A, Migranas-Ortiz R, Pérez-Miravete A, et al. Seroepidemiología de la brucelosis en México. Salud Pública Méx 1992;34:230-40.

14. Wilson SM, McNerney R, Nye O, et al. Progress toward a simplified polymerase chain reaction and its application to diagnosis tuberculosis. J Clin Microbiol. 1993;31:776-82.

15. Baily GG, Krahn JB, Drasar BS, et al. Detection of Brucella melitensis and Brucella abortus by DNA amplification. J Trop Med Hyg. 1992; 95:271-5.

16. Malorny B, Hoorfar J, Hugas M, et al. Inter-laboratory diagnostic accuracy of Salmonella specific PCR-based method. Int J Food Microbiol. 2003;89:241-9.

17. Azzimonti RJC. La concordancia entres dos tests clínicos para casos binarios: problemas y solución. ABCL. 2005;39:435-44.

18. López-Merino A, Contreras-Rodríguez A. Agentes Infecciosos: Brucella. Rev Latinoam Microbiol. 2006;48:146-53.

19. Cevallos-Falquez O, Carranza-Patiño M, Saucedo-Aguiar S, et al. Diagnóstico serológico (Rosa de Bengala) y molecular (PCR) de brucelosis en humano. Ciencia y Tecnología. 2010;3:27-32.

20. Navarro E, Casao MA, Solera J. Diagnosis of human brucellosis using PCR. Expert Rev Mol Diagn. 2004;4:115-23.

21. Nimri LF. Diagnosis of recent and relapsed cases of human brucellosis by PCR assay. BMC Infect Dis. 2003;3:5.

22. Ayyub M, Al-Juhani, NR, Alfi AY et al. Brucellosis and dengue fever -a co-infection or cross reativity? Biomedica. 2006;22:80-3.

23. Chanto G, Rojas N, Ching A, et al. Prevalencia de anticuerpos séricos contra la bacteria Brucella sp. en una población humana tropical. Rev Biol Trop. 2007;55:385-91.

24. Padilla RC, Montoya PY, Carrillo PC. Estandarización de una prueba de PCR para la detección de Brucella sp. Rev Perú Med Exp Salud Pública. 2003;20:102-4.

25. Gemechu MY, Gill JP, Arora AK, et al. Polymerase chain reaction (PCR) assay for rapid diagnosis and its role in prevention of human brucellosis in Punjab, India. Int J Prev Med. 2011;2:170-7.

26. Morata P, Queipo-Ortuño MI, Reguera JM, et al. Development and Evaluation of a PCR-Enzyme-Linked Immunoabsorbent Assay for Diagnosis of Human Brucellosis. J Clin Microbiol. 2003;41:144-8.

27. Elshamy M, Ahmed AI. The effects of maternal brucellosis on pregnancy outcome. J Infect Dev Ctries 2008;2:230-4.