Proceeding of Veterinary and Animal Science Days 2016, 8th- 10th June, Milan, Italy 

HAF © 2013 

Vol. III, No. 1s  ISSN: 2283-3927 

   

 

l 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Keywords 

LAMP, feline coronavirus, PCR 

 

CORRESPONDING AUTHOR 

Angelica Stranieri 

angelica.stranieri@unimi.it 

 

JOURNAL HOME PAGE 

riviste.unimi.it/index.php/haf 

A reverse transcription loop-mediated 

isothermal amplification (LAMP) assay for 

the detection of feline Coronavirus. 

 

A. Stranieria, S. Lauzia, A. Giordanoa, S. Paltrinieria 

 

 

aDepartment of Veterinary Medicine, Università degli Studi di Milano, Via 
Celoria 10, 20133 Milan, Italy 

 

Abstract 

Loop-mediated isothermal amplification (LAMP) is a molecular method that amplifies DNA under 

isothermal conditions. It relies on the use of 4 different primers recognizing 6 regions of the template 

sequence and on the use of a DNA polymerase with strand displacement activity (Notomi et al., 2000). 

The addition of two loop primers allows the reaction time to be of one hour only (Nagamine et al., 2002). 

The aim of this study was to develop a reverse transcription LAMP assay for an easy and inexpensive 

detection of feline Coronavirus (FCoV). Six primers binding the conserved 3’UTR region of the FCoV 

were designed with the Primer Explorer software. Thirty-two samples of RNA (11 feces, 8 effusions, 9 

blood samples and 4 tissues) on which a reverse transcription polymerase chain reaction (RT-PCR) for 

the 3’UTR region was performed were used. The reaction was carried out in 25µL reaction volume and 

the mixture was incubated in a thermocycler at 63°C for 1 hour followed by 10 minutes at 80°C. LAMP 

products were visualized under UV after electrophoresis migration on a 1.5% agarose gel stained with 

ethidium bromide, where they produce a ladder-like pattern if positive. Results where compared with 

those obtained on standard PCR. Sensitivity and specificity were respectively 60% and 100% on feces, 

40% and 100% on effusions, 25% and 100% on blood, and 100% and 100% on tissues. The overall sensitivity 

and specificity of this method were of 57.1% and 100%, thus limiting a clinical application of this method, 

except for tissues. 
 

References 

Notomi, T., Okayama, H., Masubuchi, H., Yonekawa, T., Watanabe, K., Amino, N., Hase, T., 2000. Loop-mediated isothermal 

amplification of DNA. Nucleic Acid Research. 28(12), e63-e63. 

Nagamine, K., Hase, T., Notomi, T., 2002. Accelerated reaction by loop-mediated isothermal amplification using loop primers. 

Molecular and Cellular Probes. 16(3), 223-229. 

 

http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en