Development and Validation of Loop-Mediated Isothermal Amplification (Lamp) Field Assay for the Detection of Brucella abortus

Abstract

Currently, a number of techniques are available for detection of Brucella abortus (B. abortus) but these techniques are costly and specialized equipment are needed. Therefore, the development of a rapid, accurate, sensitive, and cost effective technique for identification of Brucella species is of high priority. Objective: The current research study was designed to detect Brucella species more rapidly. The current study area was conducted in district Lodhran, Punjab, Pakistan. Methods: A total 100 blood samples (50 cattle and 50 buffaloes) were collected. Serum samples were screened against B. abortus antibodies using Rose Bengal plate test (RBPT). The specific gene was designed by using NCBI website and whole genome of Brucella species. The primers were designed from Gene accession number 20404. Following primers were designed F3, B3, FIP, BIP, LF, LB, B4, and B5. The LAMP technique for BSCP31 gene was developed by using many concentrations of components and conditions. Results: The development and validation of LAMP assay for detection of B. abortus from bovine blood in the present study proved helpful in early detection of said pathogen in animal and humans. Conclusions: This study will be helpful in prevention and control of animal and human brucellosis in Pakistan.