BACKGROUND: The diagnosis of brucellosis is frequently difficult to establish. This is not only because clinically, the disease can mimic any infectious and noninfectious disease, but also because the established diagnostic methods are not always successful. In this study, we have tried to evaluate PCR techniques in the diagnosis of brucellosis in comparison to conventional techniques. PATIENTS AND METHODS: Fifty peripheral blood samples from the following groups were collected: patients with brucellosis (17); patients with febrile illnesses due to factors other than brucella etiology (19); symptomatic occupationally exposed persons (9); and healthy volunteers (5). The last three groups were considered controls. Among the 17 Brucella samples, only 14 were obtained before treatment was begun. The samples were tested by serology, using the standard tube agglutination method (STA), blood culture using Bactec machines, and PCR using primer pair to amplify a 223-bp region within a gene coding for a 31-kD Brucella antigen. Diagnosis of brucellosis was based on compatible clinical picture in addition to positive blood culture and/or positive serology. RESULTS: Of the 17 blood samples from patients with brucellosis, eight were culture positive for Brucella species, and all showed high titer antibrucella antibodies. Only 14 of them were positive by PCR, and these were the samples submitted before initiation of therapy, representing 100% sensitivity. Among the 33 controls, blood culture was negative for Brucella in all of them, while one sample showed high-titer antibrucella antibodies. The latter was from the febrile illnesses group. PCR-based assay was able to detect four bands in the controls, all of which were from the occupationally exposed asymptomatic group. CONCLUSION: In view of the several advantages of PCR over the conventional methods for the diagnosis of brucellosis, such as speed, safety, high sensitivity and specificity, the technique might be considered for laboratory diagnosis of brucellosis. However, for the evaluation of asymptomatic highly exposed persons, PCR might be considered complementary to the traditional methods and followed up by serology and/or culture.
BACKGROUND: The diagnosis of brucellosis is frequently difficult to establish. This is not only because clinically, the disease can mimic any infectious and noninfectious disease, but also because the established diagnostic methods are not always successful. In this study, we have tried to evaluate PCR techniques in the diagnosis of brucellosis in comparison to conventional techniques. PATIENTS AND METHODS: Fifty peripheral blood samples from the following groups were collected: patients with brucellosis (17); patients with febrile illnesses due to factors other than brucella etiology (19); symptomatic occupationally exposed persons (9); and healthy volunteers (5). The last three groups were considered controls. Among the 17 Brucella samples, only 14 were obtained before treatment was begun. The samples were tested by serology, using the standard tube agglutination method (STA), blood culture using Bactec machines, and PCR using primer pair to amplify a 223-bp region within a gene coding for a 31-kD Brucella antigen. Diagnosis of brucellosis was based on compatible clinical picture in addition to positive blood culture and/or positive serology. RESULTS: Of the 17 blood samples from patients with brucellosis, eight were culture positive for Brucella species, and all showed high titer antibrucella antibodies. Only 14 of them were positive by PCR, and these were the samples submitted before initiation of therapy, representing 100% sensitivity. Among the 33 controls, blood culture was negative for Brucella in all of them, while one sample showed high-titer antibrucella antibodies. The latter was from the febrile illnesses group. PCR-based assay was able to detect four bands in the controls, all of which were from the occupationally exposed asymptomatic group. CONCLUSION: In view of the several advantages of PCR over the conventional methods for the diagnosis of brucellosis, such as speed, safety, high sensitivity and specificity, the technique might be considered for laboratory diagnosis of brucellosis. However, for the evaluation of asymptomatic highly exposed persons, PCR might be considered complementary to the traditional methods and followed up by serology and/or culture.
Authors: Andrew J Bouley; Holly M Biggs; Robyn A Stoddard; Anne B Morrissey; John A Bartlett; Isaac A Afwamba; Venance P Maro; Grace D Kinabo; Wilbrod Saganda; Sarah Cleaveland; John A Crump Journal: Am J Trop Med Hyg Date: 2012-10-22 Impact factor: 2.345