Microplate-based DNA hybridization assays for detection of human retroviral gene sequences

Abstract

Nonisotopic, microwell-based DNA hybridization assays for the specific detection of human immunodeficiency virus type 1 (HIV-1) gag, human T-cell lymphotropic virus type I (HTLV-I) pol, and HTLV-II pol DNA sequences were evaluated. The performances of these detection kits (Gene Detective enzyme oligonucleotide assays; Cellular Products, Inc., Buffalo, N.Y.) were assessed by using clinical samples whose infection status were established by amplification by PCR and then liquid hybridization detection by using virus-specific probes. Peripheral blood mononuclear cell lysates from 59 HIV-1-, 35 HTLV-I-, and 19 HTLV-II-infected individuals and from 15 healthy blood donors were used as substrates for PCR amplification. The results of the study demonstrated a clinical sensitivity of 100%k. In addition, the enzyme oligonucleotide assays were able to detect 1 to 10 proviral copies subsequent to PCR amplification, indicating an analytical sensitivity comparable to that of liquid hybridization.

Nascent human immunodeficiency virus type 1 reverse transcription occurs within an enveloped particle

“…The data presented above have several clinical implications, one of which is that in vivo HIV-1 strong-stop DNA copies may be in greater abundance than HIV-1 gag, pol, or env sequences, which are the usual targets of HIV-1 detection (1,8,21,40). Hence, viral detection strategies directed toward strong-stop DNA may prove to be more sensitive.…”

Prevalence of human herpesvirus 8 DNA sequences in several patient populations

“…DNA was organically extracted from sera and plasma. DNA from PBMs and semen was extracted with phenol-chloroform as previously outlined (17) or by a rapid lysis method (8). Fresh skin tissues were treated with proteinase K, and then the DNA was organically extracted.…”

Microbiological, immunological and molecular methods suitable for commercial detection and quantification of the blackleg pathogen, Erwinia carotovora subsp. atroseptica, on seed potato tubers: a review

“…Evaluation in five different laboratories using similar DNA extraction procedures, thermocycler, microtitre plate washer, and incubator has confirmed its sensitivity, which ranged from 10' to lo3 E. c. atroseptica cells mI-' depending on E. c. atroseptica strains, potato cultivars/stocks, and its speed and ease of use (Frkchon et al, 1998, in preparation). This DNA revelation system is already widely used in medical and plant viral diagnostics (Dyster et al, 1994;Hataya et al, 1994). Several variants of the procedure are under development and should be available soon.…”