Specific Detection of Shedding and Latency of Bovine Herpesvirus 1 and 5 using a Nested Polymerase Chain Reaction

Abstract

A sensitive method for simultaneously detecting and discriminating between bovine herpesviruses types 1 and 5 (BHV-1 and BHV-5) was developed using a nested polymerase chain reaction (PCR) technique. Following amplification using type-common primers derived from gC sequences, amplification using type-specific nesting primers produced different-sized bands specific to the corresponding types, as demonstrated by blot hybridization. Less than 0.1 plaque-forming units (PFU) of each virus and 75 fg or less of viral DNA were routinely detected. The PCR technique amplified correct product from 4 BHV-5 isolates and from 48 BHV-1 isolates, all from the United States, and did not amplify heterologous herpesviruses. The PCR technique was more sensitive than virus isolation in detection of BHV-1 or BHV-5 in nasal secretions from experimentally and naturally infected calves, and it detected BHV-1 or BHV-5 in trigeminal ganglia from these calves.

Dexamethasone-induced reactivation of bovine herpesvirus type 5 latent infection in experimentally infected rabbits results in a broader distribution of latent viral DNA in the brain

“…Therefore, oronasal infection results in the establishment of latency mainly in trigeminal ganglia (TG) and genital infection ensues, with colonization of sacral ganglia with latent viral DNA (2,(9)(10)(11)(12). However, other possible neural and nonneural sites of latent infection by, or persistence of, these viruses have also been described (13)(14)(15).…”

“…The major sites of latent infection by human (i.e., herpes simplex virus 1) and animal α-herpesviruses (BHV-5, BHV-1, pseudorabies virus) are the sensory nerve ganglia innervating the site of primary viral replication (2,(9)(10)(11). Therefore, oronasal infection results in the establishment of latency mainly in trigeminal ganglia (TG) and genital infection ensues, with colonization of sacral ganglia with latent viral DNA (2,(9)(10)(11)(12).…”

Virologic Investigations of Free-Living European Bison (Bison Bonasus) From the Bialowieza Primeval Forest, Poland

“…For BoHV-1 specific nested PCR, two primer sets amplifying a 653 bp and a final 274 bp fragment were used as described by Ashbaugh et al (1997). The PCR mixes (50 l) contained 1-2 g of cellular DNA, 0.4 M of each primer, 0.2 mM desoxyribonucleotide triphosphates (NTPs) (Applied Biosystems, Weiterstadt, Germany), 1.5 U Taq polymerase (Quiagen, Hilden, Germany), reaction buffer, and Q-solution (Quiagen).…”

Índices reprodutivos e características de desempenho em bovinos de corte infectados e não infectados pelo Herpesvírus bovino tipo 1 (HVB-1).

“…O contato direto é a principal forma de transmissão entre a fonte de infecção e o bovino susceptível, pois o HVB-1 é excretado pelas secreções respiratórias, oculares, genitais e sêmen de animais infectados (LEMAIRE et al, 1994). Os bovinos submetidos à condições estressantes, apresentam resistência imunológica reduzida, possibilitando a reativação e liberação viral do HVB-1, assegurando a permanência da infecção no plantel (ACKERMANN et al, 1982;TIKOO et al, 1995;ENGELS e ACKERMANN, 1996;KAASHOEK et al, 1996e ASHBAUGH et al, 1997 et al ( ), LEMAIRE et al (1994 e VAN DER MAATEN (1995), pois um rebanho endemicamente infectado pelo HVB-1 não apresenta elevada morbidade e mortalidade ( VAN OIRSCHOT, 1998b).…”

“…O animal portador latente do HVB -1 pode sofrer reativação viral com ou sem eliminação do vírus e uma vez infectado, será portador por toda sua vida (ACKERMANN et al, 1982;KAASHOEK et al, 1996e ASHBAUGH et al, 1997.…”