One-step nested PCR for detection of 2 LTR circles in PBMCs of HIV-1 infected patients with no detectable plasma HIV RNA.
A highly sensitive nested PCR was carried out in order to detect 2 LTR circles as a marker of recent and ongoing viral replication in HIV-1 infected patients with HIV plasma RNA undetectable. This "in house" two-step nested PCR is very sensitive, but it is not feasible for routine tests and presents a high risk of contamination. In order to reduce the time of reactions and crossover contamination, the possibility was explored to carry out a single step nested PCR, in which the two successive amplification rounds are carried out in the same tube. This single step nested PCR has the same sensitivity of the two-step nested, is easy to conduct and requires a short time of reaction. The two different PCR methods were compared and the clinical use of monitoring 2 LTR DNA circles in HIV-1 infected patients with undetectable plasma viral load is discussed.
Lusi
E A
EA
Department of Clinical Immunology and Allergy, University of Rome la Sapienza, via Portunese 332, Rome, Italy. alusi@katamail.com
Guarascio
P
P
Presutti
C
C
Villani
R
R
Pellicelli
A
A
Soccorsi
F
F
D015415:Biomarkers; D015658:HIV Infections / Q000821:virology*; D016325:HIV Long Terminal Repeat / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D012367:RNA, Viral / Q000097:blood*; D012680:Sensitivity and Specificity
D015415:Biomarkers; D012367:RNA, Viral