Detection and Differentiation of Primate α-herpesviruses by PCR

Abstract

Abstract.simplex virus types 1 and 2 [HSV1, HSV2], green monkey simian agent 8, baboon herpesvirus 2 [HVP2], and macaque B virus [BV]) was developed utilizing the polymerase chain reaction (PCR). PCR primers were located in conserved regions of the gene encoding the glycoprotein B, which flanks an intervening region that is highly divergent among the 5 viruses. Amplified PCR products from the 5 viruses were readily differentiated by their unique restriction enzyme digestion patterns. No variation in digestion patterns was noted among strains of HSV1, HSV2, or HVP2. One clinical isolate of BV exhibited variation in a single restriction site, but its overall restriction pattern remained typical of BV. This method (PCR/RFLP) allowed the presence of herpesvirus DNA in clinical swabs from primates to be readily detected and the virus unambiguously identified.

Pathogenicity of Different Baboon Herpesvirus Papio 2 Isolates Is Characterized by either Extreme Neurovirulence or Complete Apathogenicity

“…For many years the baboon virus HVP2 was assumed to be identical to SA8 (9,11,12,17). However, more recent studies show that, while very closely related to SA8, baboon isolates represent a distinct virus (1,7). Similar to the case for BV in macaques, more than 90% of adult baboons show serological evidence of infection with HVP2, and clinically evident disease due to HVP2 is uncommon (6,27).…”

“…Swab samples were processed and inoculated onto Vero or DBG3 cells as described previously (2,6). All suspected HVP2 isolates were confirmed to be HVP2 by PCR and sequencing as described previously (1,2). Origins and properties of HVP2 strains used in this study are summarized in Table 1.…”

Differential detection of B virus and rhesus cytomegalovirus in rhesus macaques FN1

“…Comparative studies of HSV detection have demonstrated that virus isolation lacks the sensitivity of PCR for the detection of virus shedding (Coyle et al, 1999;Madhavan et al, 1999;Wald et al, 1997). Traditional PCR assays for B virus detection have been described (Black & Eberle, 1997;Hirano et al, 2000;Scinicariello et al, 1993;Slomka et al, 1993). This study represents the first description of a real-time, quantitative PCR assay for B virus and its use for detection of B virus shedding from mucosal sites in macaques.…”

“…Although B virus isolation is generally considered the standard for virus detection, studies with other viruses indicate that virus isolation is a less sensitive method than other techniques such as PCR (Cone et al, 1994;Wald et al, 1995). Nonquantitative PCR techniques have been described for B virus (Black & Eberle, 1997;Scinicariello et al, 1993;Slomka et al, 1993;Weigler et al, 1995). A sensitive, quantitative PCR-based assay to determine the presence of B virus DNA would help assess both the frequency and the titre of shed viral DNA.…”

One-Step PCR To Distinguish B Virus from Related Primate Alphaherpesviruses

“…Several two-step PCR methods, which make use of restriction fragment length polymorphism (RFLP) after PCR amplification, have been developed to detect and identify BV (1,16,17,18). As previously reported, we established a PCR method to amplify a DNA segment of the glycoprotein G (gG) gene (US4 gene) from a BV strain isolated from a rhesus macaque by adding betaine (1-carboxy-N,N,N-trimethylmethanammonium inner salt) to the PCR mixture (9).…”