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The plaque reduction neutralization test is used to quantify the titer of neutralizing antibody for a virus. [1] [2] The serum sample or solution of antibody to be tested is diluted and mixed with a viral suspension. This is incubated to allow the antibody to react with the virus. This is poured over a confluent monolayer of host cells.
Surrogate virus neutralization test (sVNT) results were reported as mean inhibition rate (%). Up to day 28 (before 2nd vaccination), inhibition rates of all groups were below cut-off value of 30%. At day 35, mean inhibition of groups Nanocovax 25 mcg, 50 mcg, 75 mcg and placebo were 58.5% , 63.8% ,70.2%, and 11.1% , respectively.
This technique makes many copies of the virus genome using virus-specific probes. Variations of PCR such as nested reverse transcriptase PCR and real time PCR can also be used to determine viral loads in patient serum. This is often used to monitor treatment success in HIV cases.
However, serum therapy is today still used as the first line of defence during an outbreak as it can relatively quickly obtained. [19] [20] Serum therapy was shown to reduce mortality in patients during the 2009 swine flu pandemic [21] and the Western African Ebola virus epidemic. [22] It is also being tested as possible treatment for COVID-19.
Neutralization assays assess whether sample antibodies prevent viral infection in test cells. [67] These tests sample blood, plasma or serum. The test cultures cells that allow viral reproduction (e.g., Vero E6 cells). By varying antibody concentrations, researchers can visualize and quantify how many test antibodies block virus replication. [78]
A general procedure for HA is as follows, a serial dilution of virus is prepared across the rows in a U or V- bottom shaped 96-well microtiter plate. [5] The most concentrated sample in the first well is often diluted to be 1/5x of the stock, and subsequent wells are typically two-fold dilutions (1/10, 1/20, 1/40, etc.).
In blood grouping, the patient's serum is tested against RBCs of known blood groups and also the patient's RBCs are tested against known serum types. In this way the patient's blood group is confirmed from both RBCs and serum. A direct Coombs test is also done on the patient's blood sample in case there are any confounding antibodies.
Concentration and sterility of the serum were checked carefully, and the serum was filtered many times. Protein content was decreased in order to use the serum for humans. After the blood sampling, the horses could rest for three to four weeks and received extra food to recover the blood loss.