title: New closed tube loop mediated isothermal amplification assay for prevention of product cross-contamination.
abstract: Loop mediated isothermal amplification (LAMP) assay, a promising diagnostic test, has been developed for detection of different pathogens of human as well as animals. Various positive points support its use as a field level test but the major problem is product cross contamination leading to false positive results. Different methods were adopted by various researchers to control this false positive amplification due to cross contamination but all have their own advantages and disadvantages. A new closed tube LAMP assay based on agar dye capsule was developed in the present study and this technique has some advantages over the other closed tube technique.•Agar at the concentration of 1.5% was used to sandwich SYBR green dye I with the aid of intradermal syringe. This agar dye capsule was placed over the LAMP reaction mixture before it was amplified.•To eliminate the hazardous nature of Ultra Violet (UV) light during result visualization of LAMP products, the present study demonstrates the use of Light Emitting Diode (LED) lights for result visualization.•LAMP was carried out for Brucella species detection using this modified techniques yielding good results without any cross contamination and LED showed similar fluorescence compared to UV.
authors.affiliation: Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Bareilly 243122, India.
authors.forename: K
authors.initials: K
authors.lastname: Karthik
authors.affiliation: Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Bareilly 243122, India.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Rathore
authors.affiliation: Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Bareilly 243122, India.
authors.forename: Prasad
authors.initials: P
authors.lastname: Thomas
authors.affiliation: Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Bareilly 243122, India.
authors.forename: T R
authors.initials: TR
authors.lastname: Arun
authors.affiliation: Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Bareilly 243122, India.
authors.forename: K N
authors.initials: KN
authors.lastname: Viswas
authors.affiliation: Division of Pathology, Indian Veterinary Research Institute, Bareilly 243122, India.
authors.forename: Kuldeep
authors.initials: K
authors.lastname: Dhama
authors.affiliation: Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Bareilly 243122, India.
authors.forename: R K
authors.initials: RK
authors.lastname: Agarwal
keywords: Agar dye capsule; Brucella; Contamination; LAMP; LED; SYBR green