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2 "Davoud Afshar"
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A Novel PCR Assay for Detecting Brucella abortus and Brucella melitensis
Saeed Alamian, Majid Esmaelizad, Taghi Zahraei, Afshar Etemadi, Mohsen Mohammadi, Davoud Afshar, Soheila Ghaderi
Osong Public Health Res Perspect. 2017;8(1):65-70.   Published online February 28, 2017
  • 4,824 View
  • 70 Download
  • 8 Crossref
AbstractAbstract PDF

Brucellosis is a major zoonotic disease that poses a significant public health threat worldwide. The classical bacteriological detection process used to identify Brucella spp. is difficult and time-consuming. This study aimed to develop a novel molecular assay for detecting brucellosis.


All complete sequences of chromosome 1 with 2.1-Mbp lengths were compared among all available Brucella sequences. A unique repeat sequence (URS) locus on chromosome 1 could differentiate Brucella abortus from Brucella melitensis. A primer set was designed to flank the unique locus. A total of 136 lymph nodes and blood samples were evaluated and classified by the URS-polymerase chain reaction (PCR) method in 2013–2014.


Biochemical tests and bacteriophage typing as the golden standard indicated that all Brucella spp. isolates were B. melitensis biovar 1 and B. abortus biovar 3. The PCR results were the same as the bacteriological method for detecting Brucella spp. The sensitivity and specificity of the URS-PCR method make it suitable for detecting B. abortus and B. melitensis.


Quick detection of B. abortus and B. melitensis can provide the most effective strategies for control of these bacteria. The advantage of this method over other presented methods is that both B. abortus and B. melitensis are detectable in a single test tube. Furthermore, this method covered 100% of all B. melitensis and B. abortus biotypes. The development of this URS-PCR method is the first step toward the development of a novel kit for the molecular identification of B. abortus and B. melitensis.


Citations to this article as recorded by  
  • Development of a simplified and cost-effective sample preparation method for genotyping of human papillomavirus by next-generation sequencing
    Rungrat Jitvaropas, Ukrit Thongpoom, Vorthon Sawaswong, Kritsada Khongnomnan, Witthaya Poomipak, Kesmanee Praianantathavorn, Pornjarim Nilyanimit, Yong Poovorawan, Sunchai Payungporn
    Archives of Virology.2023;[Epub]     CrossRef
  • Bovine brucellosis – a comprehensive review
    Sandip Kumar Khurana, Anju Sehrawat, Ruchi Tiwari, Minakshi Prasad, Baldev Gulati, Muhammad Zubair Shabbir, Rajesh Chhabra, Kumaragurubaran Karthik, Shailesh Kumar Patel, Mamta Pathak, Mohd. Iqbal Yatoo, Vivek Kumar Gupta, Kuldeep Dhama, Ranjit Sah, Wanpe
    Veterinary Quarterly.2021; 41(1): 61.     CrossRef
  • Survey of Zoonotic Bacterial Pathogens in Native Foxes in Central Chile: First Record of Brucella canis Exposure
    Nicolás Galarce, Sebastián de la Fuente, Beatriz Escobar, Phillip Dettleff, Pedro Abalos, Juan Carlos Hormazábal, Roberto Flores, Nicole Sallaberry-Pincheira, Víctor Martínez
    Animals.2021; 11(7): 1980.     CrossRef
  • Development and validation of immunoassay for whole cell detection of Brucella abortus and Brucella melitensis
    Richa Hans, Pranjal Kumar Yadav, Pushpendra Kumar Sharma, Mannan Boopathi, Duraipandian Thavaselvam
    Scientific Reports.2020;[Epub]     CrossRef
  • Laboratory Diagnostic Procedures for Human Brucellosis: An Overview of Existing Approaches
    Afshar Etemadi, Rezvan Moniri, Heinrich Neubauer, Yasaman Dasteh Goli, Saeed Alamian
    Jundishapur Journal of Microbiology.2019;[Epub]     CrossRef
  • Comparison of PCR-RFLP and PFGE for determining the clonality of Brucella isolates from human and livestock specimens
    Nasrin Bahmani, Reza Mirnejad, Mohammad Reza Arabestani, Parviz Mohajerie, Seyed Hamid Hashemi, Manoochehr Karami, Mohammad Yousef Alikhani
    Saudi Journal of Biological Sciences.2019; 26(2): 256.     CrossRef
  • Designing an immunosensor for detection of Brucella abortus based on coloured silica nanoparticles
    Arash Shams, Bahareh Rahimian Zarif, Mojtaba Salouti, Reza Shapouri, Sako Mirzaii
    Artificial Cells, Nanomedicine, and Biotechnology.2019; 47(1): 2562.     CrossRef
  • Identification of Brucella genus and eight Brucella species by Luminex bead-based suspension array
    Tina S. Lusk Pfefer, Ruth Timme, Julie A. Kase
    Food Microbiology.2018; 70: 113.     CrossRef
Rapid Molecular Approach for Simultaneous Detection of Salmonella spp., Shigella spp., and Vibrio cholera
Reza Ranjbar, Ali Naghoni, Davoud Afshar, Farhad Nikkhahi, Mohsen Mohammadi
Osong Public Health Res Perspect. 2016;7(6):373-377.   Published online December 31, 2016
  • 3,578 View
  • 25 Download
  • 10 Crossref
AbstractAbstract PDF
Gastrointestinal tract infection is still one of the serious public health problems in many geographic areas and is endemic in most countries including Iran. Early detection of the gastrointestinal tract pathogens can be extremely important. The aim of the current study was to apply a shortened time-multiplex polymerase chain reaction (PCR) for rapid and simultaneous detection of Salmonella spp., Shigella spp., and Vibrio cholera.
The standard and clinical strains of Salmonella spp., Shigella spp., and V. cholerae were used in the assay. Multiplex PCR was performed and optimized based on amplification of invA, putative integrase, and ompW genes for detecting Salmonella spp., Shigella spp., and V. cholerae, respectively. The specificity of the assay was evaluated by testing 12 different bacterial species.
Only Salmonella spp., Shigella spp., and V. cholerae strains had positive results when subjected to the assay using multiplex PCR. The assay showed a high sensitivity, and no amplification products were observed in multiplex PCR with any of the other microorganisms.
Our study indicated that the invA, putative integrase, and ompW-based multiplex PCR assay appears to be an efficient method for rapid and simultaneous detection of Salmonella spp., Shigella spp., and V. cholerae.


Citations to this article as recorded by  
  • Development and Validation of an Efficient Multiplex PCR Assay for Simultaneous Detection of Six Common Foodborne Pathogens and Hygiene Indicators
    Natharin Ngamwongsatit, Soraya Chaturongakul, Ratchaneewan Aunpad
    Foodborne Pathogens and Disease.2023; 20(6): 222.     CrossRef
  • Rapid and multiplexed quantification of Salmonella, Escherichia coli O157:H7, and Shigella flexneri in ground beef using flow cytometry
    Ziquan Wang, Qian Xu, Siyuan Liu, Yingying Liu, Ying Gao, Meng Wang, Ling Zhang, Haiyan Chang, Qiang Wei, Zhiwei Sui
    Talanta.2022; 238: 123005.     CrossRef
  • Development of multiplex real-time quantitative PCR for simultaneous detection of Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, and Mycoplasma genitalium in infertile women
    Sara Sadeqi, Farhad Nikkhahi, Amir Javadi, Sonia Eskandarion, Seyed Mahmoud Amin Marashi
    Indian Journal of Medical Microbiology.2022; 40(2): 231.     CrossRef
  • Multiple fluorescent saltatory rolling circle amplification (SRCA) for simultaneous and sensitive detection of Salmonella spp. and Shigella spp. in food
    Wei Guo, Qian Yang, Jie Liu, Xiuling Chen, Yunzhe Zhang, Wei Zhang
    LWT.2022; 168: 113875.     CrossRef
  • Campylobacter Species in the Middle East
    Daryoush Babazadeh, Reza Ranjbar
    Journal of Veterinary Physiology and Pathology.2022; 1(1): 1.     CrossRef
  • Development of rapid gold nanoparticles based lateral flow assays for simultaneous detection of Shigella and Salmonella genera
    Mohammad Lukman Yahaya, Nor Dyana Zakaria, Rahmah Noordin, Khairunisak Abdul Razak
    Biotechnology and Applied Biochemistry.2021; 68(5): 1095.     CrossRef
  • Ultrasensitive pathogen detection with a rolling circle amplification-empowered multiplex electrochemical DNA sensor
    Cheryl S.Y. Yeap, Thanyarat Chaibun, Su Yin Lee, Bin Zhao, Yuan Jan, Chan La-o-vorakiat, Werasak Surareungchai, Shiping Song, Benchaporn Lertanantawong
    Chemical Communications.2021; 57(91): 12155.     CrossRef
  • Taqman hydrolysis probe application for Escherichia coli, Salmonella enterica, and Vibrio cholerae detection in surface and drinking water
    Ahmed K. A. El-Sayed, Mohamed I. Abou-Dobara, Camelia A. Abdel-Malak, Amira A. E. El-Badaly
    Journal of Water, Sanitation and Hygiene for Devel.2019; 9(3): 492.     CrossRef
  • Pathways of healthcare and antibiotics use following reported gastrointestinal illness: a cross-sectional study in rural Anhui, China
    Xing Rong Shen, Maomao Xie, Jing Chai, Rui Feng, Jing Cheng, Rong Liu, Paul Kadetz, DeBin Wang
    BMJ Open.2019; 9(8): e030986.     CrossRef
  • DNA Microarray for Rapid Detection and Identification of Food and Water Borne Bacteria: From Dry to Wet Lab
    Reza Ranjbar, Payam Behzadi, Ali Najafi, Raheleh Roudi
    The Open Microbiology Journal.2017; 11(1): 330.     CrossRef

PHRP : Osong Public Health and Research Perspectives