Detection of herpes simplex virus-1 by nested PCR. An experimental model.
Nested polymerase chain reaction (nested PCR) was performed using a reaction mix batch-prepared and kept frozen in single reaction tubes at -20 degrees C until use. Twenty-one New Zealand white rabbits were infected with herpes simplex virus type 1 (HSV-1). Eleven animals were killed on day seven and the other ten were sacrificed on day 21. Viral culture and nested PCR was used to determine the presence of HSV-1 in samples from the tongue, HSV-1 was detected in 90.47% of the animals; in 84.21% by nested PCR and in 52.63% by culture. Nested PCR assay had greater sensitivity than culture in animals sacrificed on day seven with significative difference (p < 0.05). Higher sensitivity and faster results were obtained with this method, so we found it reliable and useful in the setting of a clinical laboratory dealing with diagnosis of herpes virus infections.
Llorente
J L
JL
ENT Department, Hospital Central de Asturias Universidad de Oviedo, Spain.
Hidalgo
F I
FI
Melón
S
S
de Oña
M
M
Carreño
M
M
Suárez
C
C
D000818:Animals; D004279:DNA, Viral / Q000032:analysis*; D004195:Disease Models, Animal; D005830:Geniculate Ganglion / Q000821:virology; D006561:Herpes Simplex / Q000175:diagnosis / Q000821:virology*; D008526:Medulla Oblongata / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018139:Simplexvirus / Q000235:genetics*; D013997:Time Factors; D014059:Tongue / Q000821:virology; D012668:Trigeminal Ganglion / Q000821:virology; D019562:Viral Load
D004279:DNA, Viral