Capture of enveloped viruses using polymer tentacles containing magnetic latex particles.
A new rapid and efficient method for enveloped viruses' capture, purification and concentration has been developed. The approach is based on the use of functionalized magnetic latex particles as a carrier for capturing of the biological samples. After the capturing step, the magnetic particles are rapidly separated from the supernatant via simple permanent magnet. The nucleic acids are then directly extracted from the captured viruses on the magnetic particles using a commercial kit. The concentration methodology of viruses was first optimized for low-concentrated samples without loss of viral activity and infectivity (detected by HIV-1 p24 antigen production assay). Sensitivity of the capture assay was determined using PCR or RT-PCR method and the detection limit was found to be 10 and 16 copies for HGV and HBV respectively. The capture assay has been further used in the subtraction hybridization technique for identification of differentially expressed sequences. Finally, the lack of the tester and driver samples from the same patient by "viral positivation" of a negative serum has been circumvented using viral capture and transfer.
Arkhis
Ahmed
A
Unité Mixte CNRS-BioMérieux, ENS de Lyon, 46 allée d'Italie, 69364 Lyon Cedex 07, France.
Elaissari
Abdelhamid
A
Delair
Thierry
T
Verrier
Bernard
B
Mandrand
Bernard
B
D000596:Amino Acids / Q000032:analysis; D000956:Antigens, Viral / Q000032:analysis; D002264:Carboxylic Acids / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D029407:GB virus C / Q000302:isolation & purification / Q000378:metabolism; D006515:Hepatitis B virus / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D008280:Magnetics; D008863:Microspheres; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D012367:RNA, Viral / Q000032:analysis; D018401:Sample Size; D012680:Sensitivity and Specificity; D014779:Virus Replication; D014780:Viruses / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
D000596:Amino Acids; D000956:Antigens, Viral; D002264:Carboxylic Acids; D004279:DNA, Viral; D011108:Polymers; D012367:RNA, Viral