Translation of a laboratory-validated equine herpesvirus-1 specific real-time PCR assay into an insulated isothermal polymerase chain reaction (iiPCR) assay for point-of-need diagnosis using POCKIT™ nucleic acid analyzer

Abstract : Equine herpesvirus myeloencephalopathy (EHM), a major problem for the equine industry in the United States, is caused by equine herpesvirus-1 (EHV-1). In addition, EHV-1 is associated with upper respiratory disease, abortion, and chorioretinal lesions in horses. Here we describe the development and evaluation of an inexpensive, user-friendly insulated isothermal PCR (iiPCR) method targeting open reading 30 (ORF30) to detect both neuropathogenic and non-neuropathogenic strains on the field-deployable POCKIT™ device for point-of-need detection of EHV-1. The analytical sensitivity of the EHV-1 iiPCR assay was 13 genome equivalents per reaction. The assay did not cross react with ten non-target equine viral pathogens. Performance of the EHV-1 iiPCR assay was compared to two previously described real-time PCR (qPCR) assays in two laboratories by using 104 archived clinical samples. All 53 qPCR-positive and 46 of the 51 qPCR-negative samples tested positive and negative, respectively, by the iiPCR. The agreement between the two assays was 95.19% (confidence interval 90.48-99.90%) with a kappa value of 0.90. In conclusion, the newly developed EHV-1 iiPCR assay is robust to provide specificity and sensitivity comparable to qPCR assays for the detection of EHV-1 nucleic acid in clinical specimens.
Type de document :
Article dans une revue
Liste complète des métadonnées

https://hal-normandie-univ.archives-ouvertes.fr/hal-02167941
Contributeur : Erika Hue <>
Soumis le : vendredi 28 juin 2019 - 12:09:11
Dernière modification le : vendredi 13 septembre 2019 - 12:03:30

Identifiants

Citation

Udeni B.R. Balasuriya, Pei-Yu Alison Lee, Yun-Long Tsai, Chuan-Fu Tsai, Yu-Han Shen, et al.. Translation of a laboratory-validated equine herpesvirus-1 specific real-time PCR assay into an insulated isothermal polymerase chain reaction (iiPCR) assay for point-of-need diagnosis using POCKIT™ nucleic acid analyzer. Journal of Virological Methods, Elsevier, 2017, 241, pp.58-63. ⟨10.1016/j.jviromet.2016.12.010⟩. ⟨hal-02167941⟩

Partager

Métriques

Consultations de la notice

21