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Iran J Med Microbiol 2007, 1(3): 47-51 |
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Hedayati H, Adli Moghaddam A, Norouzian D, Tabaraie B, Ahmadi H, Siadat S D. Improved Diagnosis of Brucella in humanserum samples by double PCR assay . Iran J Med Microbiol 2007; 1 (3) :47-51
URL: http://ijmm.ir/article-1-94-en.html
URL: http://ijmm.ir/article-1-94-en.html
Hosein Hedayati 
1, Aida Adli Moghaddam2 , Dariush Norouzian2 , Bahman Tabaraie2 , Hojat Ahmadi2 , Seyed Davar Siadat2
1, Aida Adli Moghaddam2 , Dariush Norouzian2 , Bahman Tabaraie2 , Hojat Ahmadi2 , Seyed Davar Siadat2
1- Department of Quality Control, Research and Production Complex of Karaj, Pasteur Institute of Iran , mhheda@yahoo.com
2- Department of Bacterial Vaccine and Antigen Production, Research and Production Complex of Karaj, Pasteur Institute of Iran
2- Department of Bacterial Vaccine and Antigen Production, Research and Production Complex of Karaj, Pasteur Institute of Iran
Abstract: (16107 Views)
Background and Objectives: Brucellae are gram-negative intracellular pathogens which cause zoonotic
disease in humans. Clinical manifestations of brucellosis in human are variable and often are non-specific,
and the diagnosis requires fast and accurate confirmation. Since the use of serum instead of whole-blood
samples offers several advantages for nucleic acid amplification methods, in this study we developed an
improved PCR assay for the rapid and specific laboratory diagnosis of human brucellosis directly from
serum specimens.
Material and Methods: DNA was extracted from 100 µL of serum from 30 patients with acute serologic brucellosis. The PCR reaction was carried out with Specific primers. Second PCR reaction for reamplification of the first reaction products was designed.
Results: a 223 bp conserved region on the sequence encoding the 31-KDa immunogenic outer membrane protein which is specific to the genus Brucella(BCSP31) and present in all its biovars was amplified in all serum samples.
Conclusion: For confirmation and efficient amplification of the specific target, reamplification of the first PCR products had a sharper banding patterns with high sensitivity and specificity that might be considered as a new useful method for diagnosis of human brucellosis in serum specimens.
Material and Methods: DNA was extracted from 100 µL of serum from 30 patients with acute serologic brucellosis. The PCR reaction was carried out with Specific primers. Second PCR reaction for reamplification of the first reaction products was designed.
Results: a 223 bp conserved region on the sequence encoding the 31-KDa immunogenic outer membrane protein which is specific to the genus Brucella(BCSP31) and present in all its biovars was amplified in all serum samples.
Conclusion: For confirmation and efficient amplification of the specific target, reamplification of the first PCR products had a sharper banding patterns with high sensitivity and specificity that might be considered as a new useful method for diagnosis of human brucellosis in serum specimens.
Type of Study: Original Research Article |
Subject:
Molecular Microbiology
Received: 2013/11/14 | Accepted: 2013/11/14 | ePublished: 2013/11/14
Received: 2013/11/14 | Accepted: 2013/11/14 | ePublished: 2013/11/14
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