C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/68 (2006.01) C12Q 1/48 (2006.01) C12Q 1/70 (2006.01) G01N 33/569 (2006.01) G01N 33/573 (2006.01)
Patent
CA 2221769
Disclosed is a method for quantitatively determining an antibody, contained in a biological sample, having the ability to inhibit the activity of a reverse transcriptase derived from an HIV, comprising: reacting a hybridization product, immobilized on a solid phase, of a primer consisting essentially of oligodeox- ythymine nucleotide and an RNA template consisting essentially of adenine ribopolynucleotide with a reverse transcriptase derived from HIV, to thereby bind the reverse transcriptase to the hybrid- ization product; conducting a synthesis of a DNA complementary to the RNA template from a deoxymononucleotide triphosphate in the presence of the reverse transcriptase activity-inhibiting anti- body; and measuring the amount of the synthesized DNA and quanti- tatively determining the above-mentioned antibody, based on the measured amount of the synthesized DNA. The method of the present invention can be performed with ease and with high reproducibility in classifying and quantitatively determining the antibodies against the epitopes of a viral reverse transcriptase. By the method of the present invention, not only the virus having mutated during a latent period thereof, and the properties which the virus exhibits as a result of the mutation, but also a virus having ac- quired drug resistance as a result of the administration of the drugs to the host, can be studied by quantitatively determining the antibodies. Further, it has also become possible to accurate- ly and promptly determine the changes in the infected host, such as the change in the ability to produce antibodies, which occur in accordance with the progression of illness.
Méthode de dosage d'un anticorps inhibant l'activité de la transcriptase inverse provenant du virus de l'immunodéficience humaine, qui consiste à synthétiser un acide nucléique complémentaire d'un ARN matrice sensiblement constitué d'adénine ribopolynucléotide de désoxymononucléotide-3-phosphate à l'aide d'une transcriptase inverse liée à un produit d'hybridation d'une amorce sensiblement constituée de l'oligodésoxythymine nucléotide préalablement immobilisé sur une phase solide et l'ARN matrice sus-mentionné en présence d'un anticorps inhibant l'activité de la transcriptase inverse et à déterminer l'anticorps en fonction du volume d'acide nucléique ainsi synthétisé. Cette méthode permet de classer commodément les anticorps dirigés contre les épitopes de la transcriptase inverse à un degré de reproductibilité élevé et de déterminer ainsi les anticorps. En conséquence, il est possible d'étudier un virus dans sa période latente, lequel a subi des mutations successives ou a acquis une tolérance suite à l'administration d'un médicament par l'intermédiaire du dosage des anticorps. Il est également possible de constater rapidement et avec précision les changements au niveau de l'hôte infecté concernant la progression de la maladie.
Asahi Kasei Kabushiki Kaisha
Asahi Kasei Kogyo Kabushiki Kaisha
Goudreau Gage Dubuc
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