Development of a loop-mediated isothermal amplification (LAMP) assay for molecular identification of Trueperella abortisuis isolated from pigs

Marwa F E Ahmed; Mazen Alssahen; Christoph Lämmler; Geoffrey Foster; Antonia Kreitlow; Isabel Hennig-Pauka; Madeleine Plötz; Amir Abdulmawjood

doi:10.1016/j.mcp.2022.101795

Development of a loop-mediated isothermal amplification (LAMP) assay for molecular identification of Trueperella abortisuis isolated from pigs

Mol Cell Probes. 2022 Apr:62:101795. doi: 10.1016/j.mcp.2022.101795. Epub 2022 Feb 4.

Authors

Marwa F E Ahmed¹, Mazen Alssahen², Christoph Lämmler², Geoffrey Foster³, Antonia Kreitlow⁴, Isabel Hennig-Pauka⁵, Madeleine Plötz⁴, Amir Abdulmawjood⁶

Affiliations

¹ Hygiene and Zoonoses Department, Faculty of Veterinary Medicine, Mansoura University, Elgomhoria Street 60, 35516, Mansoura, Egypt; Institute for Animal Nutrition, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, D-30173, Hannover, Germany.
² Institut für Hygiene und Infektionskrankheiten der Tiere, Justus-Liebig-Universität Gießen, Frankfurterstraße 85-91, D-35392, Gießen, Germany.
³ SRUC Veterinary Services, 10 an Lochran, Inverness Campus, Inverness, IV2 5NA, UK.
⁴ Institute of Food Quality and Food Safety, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, D-30173, Hannover, Germany.
⁵ Field Station for Epidemiology, University of Veterinary Medicine Hannover, Foundation, Büscheler Str. 9, 49456, Bakum, Germany.
⁶ Institute of Food Quality and Food Safety, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, D-30173, Hannover, Germany. Electronic address: amir.abdulmawjood@tiho-hannover.de.

PMID: 35131429
DOI: 10.1016/j.mcp.2022.101795

Abstract

The first description of Trueperella (T.) abortisuis was presented in Japan in 2009 by Azuma and colleagues. In the current study, eight T. abortisuis strains were identified by a newly developed loop-mediated isothermal amplification (LAMP) assay based on the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) encoding gene gap. Two T. abortisuis strains were obtained from prepuce of a seven-month-old boar and pooled foetal stomach contents in the United Kingdom, while the other six T. abortisuis strains were recovered from aborted foetal material of six pigs from a single farm in Germany. The developed LAMP assay showed an analytical sensitivity of 22 pg μL^-1T. abortisuis DNA. T. abortisuis DSM 19515^T and field strain T. abortisuis P504054/19/1 were directly detectable in artificially contaminated vaginal swabs up to concentrations of 980 CFU and 770 CFU per swab, respectively. There was no cross reactivity with control strains representing six species of genus Trueperella and six species of the closely related genus Arcanobacterium and Schaalia (Actinomyces) hyovaginalis. Further field research is required to determine the usefulness of the designed LAMP assay for identifying T. abortisuis isolated from pigs of various origins and from test samples directly obtained at farm level.

Keywords: LAMP assay; Pig; Trueperella abortisuis; gap.

MeSH terms

Actinomycetaceae
Animals
Arcanobacterium* / genetics
Female
Male
Molecular Diagnostic Techniques
Nucleic Acid Amplification Techniques*
RNA, Ribosomal, 16S / genetics
Sensitivity and Specificity
Swine

Substances

RNA, Ribosomal, 16S

Supplementary concepts

LAMP assay
Trueperella abortisuis