USE OF COMPETITIVE POLYMERASE CHAIN-REACTION TO DETERMINE HIV-1 LEVELS IN RESPONSE TO ANTIVIRAL TREATMENTS

被引:26
|
作者
BRUISTEN, SM
KOPPELMAN, MHGM
ROOS, MTL
LOELIGER, AE
REISS, P
BOUCHER, CAB
HUISMAN, HG
机构
[1] NETHERLANDS RED CROSS,BLOOD TRANSFUS SERV,CENT LAB,1066 CX AMSTERDAM,NETHERLANDS
[2] ACAD MED CENTRUM AMSTERDAM,NATL AIDS THERAPY EVALUAT CTR,AMSTERDAM,NETHERLANDS
关键词
COMPETITIVE POLYMERASE CHAIN REACTION; HIV-1; ANTIVIRAL TREATMENT;
D O I
10.1097/00002030-199311002-00005
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Objective: To develop a competitive polymerase chain reaction technique with which to evaluate the usefulness of HIV-1 level as a marker of response to antiviral treatment. Design: HIV-1 sequences were assessed by competitive polymerase chain reaction in four subjects participating in a double-blind study of monotherapy versus combination therapy with nucleoside analogues. Methods: We inserted a mutant construct of the HIV-1 pol sequence into a commercial vector, enabling us to generate known amounts of mutant DNA and RNA for competitive polymerase chain reaction. To measure HIV-1 DNA copies in cells, the mutant DNA fragments were allowed to compete in a 10-fold dilution series with a constant amount of nucleic acid from the subject. To measure HIV-1 RNA copies in plasma, in vitro synthesized mutant RNA was added in a 10-fold dilution series to a constant amount of subject RNA and copy DNA was synthesized. DNA and copy DNA were used as the input for nested pol polymerase chain reaction. Mutant and wild-type amplimers were discriminated by size. Results: The competitive polymerase chain reaction technique has been validated in model experiments and can be used over a broad range (at least 6 logs) of levels. Three of the four subjects showed a decline of 1 log in proviral DNA levels in cells after beginning antiviral treatment. All four showed a decline of at least 1 log in viral RNA levels in plasma, but this decline was transient in one subject. Conclusion: The HIV-1 sequence level is a useful marker in antiviral treatment studies.
引用
收藏
页码:S15 / S20
页数:6
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