ELISAs are commonly used in clinical laboratories to test for SARS antibodies. However, despite their popularity, some kits are not as sensitive or specific as others. This is where high throughput ELISA testing can make a huge difference. Not only do ELISAs increase the speed of results, but they can also be used during public health emergencies. ELISA stands for Enzyme-Linked ImmunoSorbent Assays.
These systems are based on the simple step ELISA protocol. They allow researchers to process four times as many samples in the same amount of time. The SimpleStep ELISA kit uses a 384-well format that allows for four times the number of samples processed. Another example of high throughput ELISA is the PHERAstar FSX, which measures a single wavelength per well in under a second. Additionally, these kits use BMG LABTECH's MARS data analysis software to automatically generate quantitative standard curves.
ELISA is an extremely common quantitative method for detecting a target antigen, toxin, or foreign substance. But the difficulty lies in translating its usefulness into high throughput screening. This article explores the challenges that ELISA faces in high throughput applications and recommends alternatives. And if these challenges still exist for your laboratory, you can always turn to a higher-throughput ELISA system.
A new generation of biochips designed for high throughput ELISAs is available today. These are optically flat glass plates that contain 96 wells surrounded by hydrophobic Teflon masks. Their footprint dimensions match standard microplates. The four identical 36-element arrays containing eight different antigens are placed in each well. High speed X-Y-Z robots print the arrays on the chips. With a single robot, the production rate of an array can reach twenty thousand a day.
HSV-2 rgG2 ELISA is a purified recombinant protein from baculovirus that uses native HSV antigens to inhibit gG2 antibody binding. This method distinguishes between nonspecific and specific gG2 antibody binding. 260 positive samples from the Kenya-A, South Africa, Uganda-A, Zimbabwe panels were tested for differential blocking abilities.
The results of the ELISA were compared to WB methods. A significant difference was noted between the two. WB compared to ELISA results were not always identical, and HSV-2 lysate was able to produce different inhibitory effects in different sera. Therefore, this method may be preferred for comparing two-stage ELISA results. While this method can be more sensitive and accurate, it is limited by its limited cross-reactivity.
In the current studies, the Kalon HSV-2 IgG ELISA has provided reliable results for determining HSV-2 serostatus in patients. Nevertheless, the focus of the study is on establishing an FDA-cleared method. Kalon's robustness and reliability can make it a useful replacement for the current methods in resource-constrained settings. This method may be particularly useful in sub-Saharan Africa and may have broader utility outside epidemiological research.
The HerpeSelect HSV-2 ELISA detected seroconversion in an HSV-2 primary infection earlier than WB profiles. In fact, nine of 11 samples with atypical WB profiles were HSV-2-positive by the HerpeSelect and igg-inhibition ELISA. Two samples from Kenya had high HSV IgM levels, which indicate recent exposure to the virus. After detetion, there maybe some residual substances on the ELISA plate. In order to reduce the errors caused by the residues, an ELISA washer is needed.
Although the HSV-2 IgG ELISA is the gold standard for this test, it is highly expensive and technically challenging to use. The alternative is commercial ELISAs. The Focus HerpeSelect-2 IgG ELISA is FDA-cleared, while the Kalon HSV-2 IgG ELISA is not approved and should be used in secondary research only.
The index value of the HSV-2 ELISA and WB was calculated from differential reactivity of the two lysates. The cutoff criteria for the inhibition assay was determined using the formula given above. The HSV-2 lysate was used to control for loss of binding to gG2 due to other factors than the native gG2. Moreover, the WB-positive samples had higher inhibition levels than the non-infected samples.
HerpeSelect ELISA has high sensitivities, but low specificities, particularly in subpopulations in Africa. However, Kalon had higher specificity, and the latter had higher sensitivity than HerpeSelect. These results suggest the need for type-specific serological assays for subpopulations in Africa. So, the ELISA method developed by Kalon was superior to the HerpeSelect ELISA in Kisumu, Kenya.
The focus diagnostics HSV-2 gG2 inhibition ELISA proved to be an effective supplemental tool in confirming genital herpes infection. However, the results obtained in the study were not applicable in other situations due to its low sample size. Thus, it is important to remember that the NPV and PPV values obtained from ELISA testing are dependent on the prevalence of HSV-2 in the population studied.
In the present study, researchers tested 14 food-specific IgG antibodies to assess the role of ELISA in the detection of HSV-2. The foods tested included fish, beef, soy, mushrooms, and rice. The researchers also analyzed the impact of a low level of IgG in the HSV-2 infection on the ELISA results. The results indicated the role of serological testing in prevention programs for high-risk populations.