Universität zu Köln

Wichianchot, Sakulchit, Hongsrichan, Nuttanan, Maneeruttanarungroj, Cherdsak, Pinlaor, Somchai, Iamrod, Kantapong, Purisarn, Andaman, Donthaisong, Peerawich, Karanis, Panagiotis, Nimsuphan, Burin and Rucksaken, Rucksak ORCID: 0000-0002-0833-3729 (2022). A newly developed droplet digital PCR for Ehrlichia canis detection: comparisons to conventional PCR and blood smear techniques. J. Vet. Med. Sci., 84 (6). S. 831 - 841. TOKYO: JAPAN SOC VET SCI. ISSN 1347-7439

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Abstract

Canine monocytic ehrlichiosis caused by Ehrlichia canis infection is a life-threatening vector-borne disease in dogs worldwide. Routine blood smear has very low sensitivity and cannot accurately provide a quantitative result. Conventional PCR (cPCR) and real-time PCR (qPCR) are widely used as molecular methods for E. canis detection. qPCR is quantitative but relies on standard curves of known samples. To overcome this difficulty, this study developed a new E. canis quantitative detection method, using droplet digital polymerase chain reaction (ddPCR). ddPCR was evaluated against cPCR and blood smears. PCR amplicons and genomic DNA (gDNA) from 12 microscopic positive samples were used to identify the limits of detection (LODs) in ddPCR and cPCR. Our ddPCR was assessed in 92 field samples, it was compared with cPCR and blood smears. ddPCR showed LOD=1.6 copies/reaction, or 78 times more sensitive than cPCR (LOD=126 copies/reaction), using PCR amplicons as a template, whereas both ddPCR and cPCR had equal LODs at 0.02 ng gDNA/reaction. In addition, ddPCR had 100% sensitivity and 75% specificity for E. canis detection compared to cPCR and no cross-reaction with other blood pathogens was observed. ddPCR identified more positive samples than cPCR and blood smear. ddPCR improved the overall performance of E. canis detection, with a better LOD and comparable sensitivity and specificity to cPCR. The technique might be helpful for diagnosis of E. canis in light infection, evaluating the number of E. canis and follow-up after treatment.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Wichianchot, Sakulchit UNSPECIFIED UNSPECIFIED UNSPECIFIED Hongsrichan, Nuttanan UNSPECIFIED UNSPECIFIED UNSPECIFIED Maneeruttanarungroj, Cherdsak UNSPECIFIED UNSPECIFIED UNSPECIFIED Pinlaor, Somchai UNSPECIFIED UNSPECIFIED UNSPECIFIED Iamrod, Kantapong UNSPECIFIED UNSPECIFIED UNSPECIFIED Purisarn, Andaman UNSPECIFIED UNSPECIFIED UNSPECIFIED Donthaisong, Peerawich UNSPECIFIED UNSPECIFIED UNSPECIFIED Karanis, Panagiotis UNSPECIFIED UNSPECIFIED UNSPECIFIED Nimsuphan, Burin UNSPECIFIED UNSPECIFIED UNSPECIFIED Rucksaken, Rucksak UNSPECIFIED orcid.org/0000-0002-0833-3729 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-683424
DOI:	10.1292/jvms.22-0086
Journal or Publication Title:	J. Vet. Med. Sci.
Volume:	84
Number:	6
Page Range:	S. 831 - 841
Date:	2022
Publisher:	JAPAN SOC VET SCI
Place of Publication:	TOKYO
ISSN:	1347-7439
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language POLYMERASE-CHAIN-REACTION; TICK-BORNE PATHOGENS; ANAPLASMA-PLATYS; MONOCYTIC EHRLICHIOSIS; MOLECULAR-DETECTION; BABESIA-CANIS; HEPATOZOON-CANIS; HUMAN INFECTION; STRAY DOGS; DIAGNOSIS Multiple languages Veterinary Sciences Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/68342