Cargando…

Real-Time Fluorometric Isothermal LAMP Assay for Detection of Chlamydia pecorum in Rapidly Processed Ovine Abortion Samples: A Veterinary Practitioner’s Perspective

Traditional methods of detecting Chlamydia pecorum in tissue samples such as polymerase chain reaction or cell culture are laborious and costly. We evaluated the use of a previously developed C. pecorum LAMP assay using minimally processed ovine samples. Cotyledon (n = 16), foetal liver (n = 22), fo...

Descripción completa

Detalles Bibliográficos
Autores principales: Clune, Tom, Anstey, Susan, Kasimov, Vasilli, Jacobson, Caroline, Jelocnik, Martina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8470028/
https://www.ncbi.nlm.nih.gov/pubmed/34578188
http://dx.doi.org/10.3390/pathogens10091157
_version_ 1784574093051297792
author Clune, Tom
Anstey, Susan
Kasimov, Vasilli
Jacobson, Caroline
Jelocnik, Martina
author_facet Clune, Tom
Anstey, Susan
Kasimov, Vasilli
Jacobson, Caroline
Jelocnik, Martina
author_sort Clune, Tom
collection PubMed
description Traditional methods of detecting Chlamydia pecorum in tissue samples such as polymerase chain reaction or cell culture are laborious and costly. We evaluated the use of a previously developed C. pecorum LAMP assay using minimally processed ovine samples. Cotyledon (n = 16), foetal liver (n = 22), foetal lung (n = 2), and vaginal (n = 6) swabs, in addition to cotyledon (n = 6) and foetal liver (n = 8) tissue samples, were rapidly processed and used for LAMP testing without DNA extraction. Overall, LAMP test results were highly congruent with the in-house reference qPCR, with 80.43% (37/46; 72.73% positive agreement (PA); 84.75% negative agreement (NA)) overall agreeance for swab samples, and 85.71% (12/14; 80% PA; 88.89% NA) overall agreeance for tissue samples. Out of the 11 total discrepant results, discrepancy was mainly observed in samples (n = 10) with less than 100 copies/µL C. pecorum DNA. While sensitivity could be improved, the simplicity, low cost, and accuracy of detection makes this test amenable for use at point-of-care for detecting C. pecorum in sheep.
format Online
Article
Text
id pubmed-8470028
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-84700282021-09-27 Real-Time Fluorometric Isothermal LAMP Assay for Detection of Chlamydia pecorum in Rapidly Processed Ovine Abortion Samples: A Veterinary Practitioner’s Perspective Clune, Tom Anstey, Susan Kasimov, Vasilli Jacobson, Caroline Jelocnik, Martina Pathogens Communication Traditional methods of detecting Chlamydia pecorum in tissue samples such as polymerase chain reaction or cell culture are laborious and costly. We evaluated the use of a previously developed C. pecorum LAMP assay using minimally processed ovine samples. Cotyledon (n = 16), foetal liver (n = 22), foetal lung (n = 2), and vaginal (n = 6) swabs, in addition to cotyledon (n = 6) and foetal liver (n = 8) tissue samples, were rapidly processed and used for LAMP testing without DNA extraction. Overall, LAMP test results were highly congruent with the in-house reference qPCR, with 80.43% (37/46; 72.73% positive agreement (PA); 84.75% negative agreement (NA)) overall agreeance for swab samples, and 85.71% (12/14; 80% PA; 88.89% NA) overall agreeance for tissue samples. Out of the 11 total discrepant results, discrepancy was mainly observed in samples (n = 10) with less than 100 copies/µL C. pecorum DNA. While sensitivity could be improved, the simplicity, low cost, and accuracy of detection makes this test amenable for use at point-of-care for detecting C. pecorum in sheep. MDPI 2021-09-08 /pmc/articles/PMC8470028/ /pubmed/34578188 http://dx.doi.org/10.3390/pathogens10091157 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Clune, Tom
Anstey, Susan
Kasimov, Vasilli
Jacobson, Caroline
Jelocnik, Martina
Real-Time Fluorometric Isothermal LAMP Assay for Detection of Chlamydia pecorum in Rapidly Processed Ovine Abortion Samples: A Veterinary Practitioner’s Perspective
title Real-Time Fluorometric Isothermal LAMP Assay for Detection of Chlamydia pecorum in Rapidly Processed Ovine Abortion Samples: A Veterinary Practitioner’s Perspective
title_full Real-Time Fluorometric Isothermal LAMP Assay for Detection of Chlamydia pecorum in Rapidly Processed Ovine Abortion Samples: A Veterinary Practitioner’s Perspective
title_fullStr Real-Time Fluorometric Isothermal LAMP Assay for Detection of Chlamydia pecorum in Rapidly Processed Ovine Abortion Samples: A Veterinary Practitioner’s Perspective
title_full_unstemmed Real-Time Fluorometric Isothermal LAMP Assay for Detection of Chlamydia pecorum in Rapidly Processed Ovine Abortion Samples: A Veterinary Practitioner’s Perspective
title_short Real-Time Fluorometric Isothermal LAMP Assay for Detection of Chlamydia pecorum in Rapidly Processed Ovine Abortion Samples: A Veterinary Practitioner’s Perspective
title_sort real-time fluorometric isothermal lamp assay for detection of chlamydia pecorum in rapidly processed ovine abortion samples: a veterinary practitioner’s perspective
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8470028/
https://www.ncbi.nlm.nih.gov/pubmed/34578188
http://dx.doi.org/10.3390/pathogens10091157
work_keys_str_mv AT clunetom realtimefluorometricisothermallampassayfordetectionofchlamydiapecoruminrapidlyprocessedovineabortionsamplesaveterinarypractitionersperspective
AT ansteysusan realtimefluorometricisothermallampassayfordetectionofchlamydiapecoruminrapidlyprocessedovineabortionsamplesaveterinarypractitionersperspective
AT kasimovvasilli realtimefluorometricisothermallampassayfordetectionofchlamydiapecoruminrapidlyprocessedovineabortionsamplesaveterinarypractitionersperspective
AT jacobsoncaroline realtimefluorometricisothermallampassayfordetectionofchlamydiapecoruminrapidlyprocessedovineabortionsamplesaveterinarypractitionersperspective
AT jelocnikmartina realtimefluorometricisothermallampassayfordetectionofchlamydiapecoruminrapidlyprocessedovineabortionsamplesaveterinarypractitionersperspective