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Open Access Research article

Two-stage PCR assay for detection of human brucellosis in endemic areas

Ibrahim Hassan Kamal12*, Basim Al Gashgari1, Said Salama Moselhy12, Taha Abdullah Kumosani13 and Khalid Omar Abulnaja1

Author Affiliations

1 Biochemistry Department, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia

2 Biochemistry Department, Faculty of Science, Ain Shams University, Cairo, Egypt

3 Experimental Biochemistry Unit, King Fahd Medical Research Center (KFMRC), King Abdulaziz University, Jeddah, Saudi Arabia

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BMC Infectious Diseases 2013, 13:145  doi:10.1186/1471-2334-13-145

Published: 21 March 2013

Abstract

Background

Brucellosis is a common zoonosis that can cause a severe febrile illness in humans. It constitutes a persistent health problem in many developing countries around the world. It is one of the most frequently reported diseases in Saudi Arabia and incidence is particularly high in the Central region, and around the city of Riyadh. The aim of this study was to evaluate a two-stage PCR assay for detection of human brucellosis particularly in endemic areas.

Methods

A total of 101 serum samples were collected from patients with acute febrile illness (AFI) of unknown cause from two different locations in the Western region of Saudi Arabia. The first location (Northern) is characterized by a nomadic rural population while the second (Central) is a modern urban city. All samples were subjected to DNA extraction and Brucella genus-specific PCR amplification using B4/B5 primers of the bcsp31 gene. Positive B4/B5 samples were subjected to multiplex species-specific Brucella PCR amplification.

Results

In the Northern location, 81.9% of the AFI samples were confirmed Brucella positive, while all the samples collected from the Central region proved to be Brucella negative. Samples positive for Brucella were subjected to multiplex species-specific Brucella amplification. B. abortus was detected in 10% and B. melitensis in 8% of the samples, while the majority (82%) of samples showed both B. abortus and B. melitensis. As expected, B. suis was not detected in any of the samples.

Conclusions

This study concluded that a two-stage PCR assay could be useful as a rapid diagnostic tool to allow the consideration of brucellosis as a possible cause of AFI, particularly in non-urban locations. It also recommends the collection of epidemiological data for such patients to obtain further information that may help in rapid diagnosis.