RT-PCR detection of lentiviruses in milk or mammary secretions of sheep or goats from infected flocks.
In this study we evaluated a reverse transcriptase polymerase chain reaction (RT-PCR) technique for detecting lentiviral infection in milk or mammary secretions from small ruminants. Initial observations on seven goats infected with cloned caprine arthritis-encephalitis virus (CAEV) showed that RT-PCR on milk cells is as reliable as coculture for detecting viral infection, and is quicker and simpler. With a suitable choice of redundant primers followed by a semi-nested amplification, it proved possible to detect the virus in milk samples from naturally infected French sheep (8/8) or goats (9/9), and viral sub-groups could be identified by hybridization with discriminatory probes. All seropositive animals gave positive amplifications, as did one seronegative goat from a contaminated herd, suggesting greater sensitivity for RT-PCR. None of eight goats from a long-established seronegative herd ever gave a positive RT-PCR amplification. This technique provides a simple means for rapidly identifying potentially infectious animals and for epidemiological investigations, as long as the primers are selected according to the genetic structure of the local viral population.
Leroux
C
C
Laboratoire d'immunologie et de biologie pulmonaire, INSERM CJF 93-08, université Claude-Bernard.
Lerondelle
C
C
Chastang
J
J
Mornex
J F
JF
D000818:Animals; D016088:Arthritis-Encephalitis Virus, Caprine / Q000235:genetics / Q000302:isolation & purification*; D001483:Base Sequence; D017931:DNA Primers; D005260:Female; D016679:Genome, Viral; D015511:Goat Diseases; D006041:Goats; D016086:Lentivirus / Q000235:genetics / Q000302:isolation & purification*; D016180:Lentivirus Infections / Q000175:diagnosis / Q000662:veterinary*; D008321:Mammary Glands, Animal / Q000378:metabolism / Q000821:virology*; D008892:Milk / Q000821:virology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012756:Sheep; D012757:Sheep Diseases
D017931:DNA Primers