A semiquantitative microassay for measurement of relative number of blood mononuclear cells infected with human immunodeficiency virus
A easy semiquantitative microassay was developed for the measurement of relative variety of contaminated peripheral blood mononuclear cells (PBMC) from people contaminated with human immunodeficiency virus (HIV). The assay relies on cocultivation of serially diluted PBMC of a seropositive individual with phytohemagglutinin-stimulated regular PBMC. The microassay has comparable sensitivity with the usual virus tradition methodology in detecting constructive HIV cultures.
Because the microassay makes use of solely 2-Three x 10(5) sufferers’ PBMC, the assay can be most fitted for HIV isolation from HIV-infected infants or from AIDS sufferers with extraordinarily low T-cell counts. The microassay will also be used to measure antiviral results of a drug on persistent HIV an infection in vitro. As a result of the microassay measures the relative variety of contaminated PBMC, it may be readily used for following the quantitative antiviral impact of a drug in a scientific trial.
A microassay to quantitate collagen synthesis by cells in tradition
A technique to quantitate collagen synthesis, complete protein synthesis, and DNA in 24-well tradition plates is offered. Collagen-producing cells equivalent to human intestinal clean muscle cells and dermal fibroblasts have been pulse-labeled with [3H]proline. After incubation, the plates have been heated to 90 levels C to cease isotope incorporation and sonicated to lyse the cells and an aliquot was eliminated for DNA quantitation. Service protein was added, all protein was precipitated by trichloroacetic acid, and unbound isotope was eliminated by repeated precipitations.
After incubation with purified bacterial collagenase, each the soluble 3H-labeled collagen-derived peptides and the remaining insoluble 3H-labeled noncollagen protein have been quantified. Outcomes have been expressed as the quantity of radioactivity integrated into collagen and noncollagen protein per nanogram DNA and in addition as the share of collagen synthesis per complete protein synthesized.
The benefit of this method over earlier makes an attempt to scale down the assay is that your entire assay for DNA, collagen, and non-collagen protein will be carried out in the identical nicely with none switch of fabric. This system additionally supplies a big financial savings of tradition medium, serum, development elements, and cell materials.
This research describes a dot enzyme immunoassay (Dot-EIA) for detecting hepatitis B floor antigen (HBsAg). The outcomes demonstrated that the detection stage of this assay for HBsAg was 1.5 ng/ml; no false-positive or -negative readings have been noticed. Additionally, this Dot-EIA had some benefits over normal EIA: (1) antiserum could possibly be straight and instantly certain on nitrocellulose paper set into microfiltration equipment, (2) the paper could possibly be simply washed underneath decreased strain utilizing a water aspirator, (3) all assay steps could possibly be carried out at room temperature inside 2 h, (4) the well-defined brown spots could possibly be evaluated by each visible remark and densitometric studying. The Dot-EIA reported right here could also be helpful for speedy prognosis and screening of HBsAg in serum.