Serologic and molecular diagnosis of Colorado tick fever viral infections.
Molecular and serologic methods usable for the biological diagnosis of Coltivirus infection are reported. We designed a multiplex reverse transcription-polymerase chain reaction system that allowed the simultaneous and specific amplification of three genomic segments from as little as 0.01 plaque-forming units. Another system in the S2 viral segment permitted the differential diagnosis of American and European viral isolates. We also discuss some improvements of previous ELISAs, and the results obtained with paired sera from Colorado tick fever (CTF) virus-infected individuals. Western blot analysis was developed that allowed the detection of antibodies to a 38-kD viral protein in all tested sera. It also enabled the detection of anti-CTF virus antibodies in ELISA-negative sera. Specific IgM antibodies against a synthetic viral peptide could be detected in sera at the acute stage of the infection. Together, these results should permit the diagnosis of Coltivirus infection at any stage of the pathology.
Attoui
H
H
Laboratoire de Virologie Moléculaire, Tropicale et Transfusionnele, Faculté de Médecine de Marseille, France.
Billoir
F
F
Bruey
J M
JM
de Micco
P
P
de Lamballerie
X
X
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western / Q000379:methods / Q000706:statistics & numerical data; D002460:Cell Line; D003121:Colorado Tick Fever / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D003122:Colorado tick fever virus / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D006224:Cricetinae; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000706:statistics & numerical data; D005069:Evaluation Studies as Topic; D005260:Female; D006801:Humans; D007075:Immunoglobulin M / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods* / Q000706:statistics & numerical data; D014773:Virology / Q000379:methods* / Q000706:statistics & numerical data
D000914:Antibodies, Viral; D000956:Antigens, Viral; D017931:DNA Primers; D007075:Immunoglobulin M