A sensitive haemadsorption technique based RT-PCR for concentration and detection of Newcastle disease virus from clinical samples and allantoic fluid.

Autor: Desingu PA; Avian Diseases Section, Division of Pathology, ICAR-Indian Veterinary Research Institute (IVRI), Izatnagar, Bareilly, UP 243 122 India., Singh SD; Avian Diseases Section, Division of Pathology, ICAR-Indian Veterinary Research Institute (IVRI), Izatnagar, Bareilly, UP 243 122 India., Dhama K; Avian Diseases Section, Division of Pathology, ICAR-Indian Veterinary Research Institute (IVRI), Izatnagar, Bareilly, UP 243 122 India., Vinodhkumar OR; Division of Epidemiology, ICAR-Indian Veterinary Research Institute (IVRI), Izatnagar, UP 243 122 India., Malik YS; Division of Biological Standardization, ICAR-Indian Veterinary Research Institute (IVRI), Izatnagar, UP 243 122 India.
Jazyk: angličtina
Zdroj: Virusdisease [Virusdisease] 2016 Sep; Vol. 27 (3), pp. 319-323. Date of Electronic Publication: 2016 Jun 11.
DOI: 10.1007/s13337-016-0325-9
Abstrakt: The present study describes the exploitation of haemadsorption (HAd) property of the Newcastle disease virus (NDV) for the development of a novel sensitive HAd technique based RT-PCR for detection of NDV from clinical samples of virus infected experimental birds. The NDV propagated allantoic fluid from the infected embryonated chicken eggs or supernatant of the processed clinical samples (tissue triturate, cloaca and tracheal swabs) from the experimentally infected birds were added with chicken red blood cells (RBC) to adsorb the virus on RBC's surface. The virus adsorbed RBCs were subjected to trizol method of RNA extraction and reverse transcription-polymerase chain reaction (RT-PCR) for detection of NDV. The HAd based RNA extraction showed better yield of 700-900 ng RNA and when subjected to RT-PCR detection revealed a 100 times higher sensitivity than the conventional RNA extraction and RT-PCR detection system. This could be an alternate technique which can be exploited in low NDV load situations in clinical samples.
Databáze: MEDLINE
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