Utilization of the polymerase chain reaction in the diagnosis of nuclear polyhedrosis virus infections of gypsy moth (Lymantria dispar, Lep., Lymantriidae) populations

• Published in: Journal of applied entomology (1986) Vol. 127; no. 7; pp. 405 - 412
• Main Authors: Charpentier, G, Desmarteaux, D, Bourassa, J.P, Belloncik, S, Arella, M
• Format: Journal Article
• Language: English
• Published: Berlin, Germany Blackwell Verlag GmbH 01-08-2003; Blackwell
• Subjects: Biological and medical sciences; cloning method; diagnostic techniques; disease detection; disease diagnosis; disease transmission; DNA probes; egg masses; Fundamental and applied biological sciences. Psychology; infection; insect pests; insect viruses; larvae; LdNPV genome; Lymantria dispar; Lymantria dispar multiple nucleopolyhedrovirus; natural infection; natural infections; Nuclear polyhedrosis virus; pathogen identification; PCR; Phytopathology. Animal pests. Plant and forest protection; polymerase chain reaction; Protozoa. Invertebrates; viral transmission; Canada; United States; USA; Virus; Infection; Nucleopolyhedrovirus; Polymerase chain reaction; Lymantriidae; Baculoviridae; Arthropoda; Insecta; Lepidoptera; Invertebrata; Lymantria dispar
• ISSN: 0931-2048; 1439-0418
• DOI: 10.1046/j.1439-0418.2003.00759.x

:  Three specific DNA probes were used for the detection of the nuclear polyhedrosis (NPV) virus of Lymantria dispar (LdNPV) genome. Two of these probes, H2 and H3 were obtained by classical cloning method and one (TR6) by polymerase chain reaction (PCR). These probes, used individually or in a pool in the standard slot–blot hybridizations, were able to detect 109 genome copies. By performing 35 cycles of PCR amplification before hybridization with primers specific to LdNPV genome on DNA extracted from infected larvae, the sensitivity of the hybridization technique was increased, so that as little as 10 copies of the LdNPV genome could be detected. Using these methods, L. dispar naturally infected by LdNPV were identified among field populations in Canada and in the United States near the eastern Canadian border. Using a combination of PCR and hybridization, LdNPV contamination of egg masses were also detected. By disinfecting the eggs with sodium hypochlorite prior to PCR amplification and hybridization, it was also demonstrated that transmission of viral infection in the natural populations is mainly caused by external contamination of the egg and is unlikely to occur through the transovarial route.