I would appreciate comments especially from Ketsuko, Fowlerstoad and Phage.
I just flip burgers at a hamburger joint and know nothing about anything. But I have to wonder why several patients are initially testing negative
only to retest positive. One example is found here
Something in the current RT PCR test is looking for a substance that the current reagent is not finding quickly enough. Here is what the current PCR
is looking at:
The above represent relative positions of amplicon targets on SARS-CoV ad Wuhan-CoV genome.N: nucleocapsid; ORF: open reading frame; RdRp:
RNA-dependent RNA polymerase.Numbers below amplicon are genome positions according to SARS-CoV, NC_004718.
This diagram represents what the PCR is looking for.
The PCR is looking for what 6-Carboxyfluorescein (6-FAM) attaches too. 6-FAM attaches to the Oligonucleotide RdRp_SARSr-P2 and RdRP_SARSr-P1 It is
the test used by the CDC and WHO.
6-FAM is a fluorescent dye and is membrane-impermeant, and can be loaded into cells by microinjection or scrape loading. It can be incorporated into
liposomes, and allow for the tracking of liposomes as they pass through the body. A liposome is a spherical vesicle having at least one lipid bilayer.
A microfluidic based mixing of lipids, helper lipid DSPC (1), PEG-lipid (2), ionizable amino lipid DLin-MC3-DMA
and cholesterol(4), diluted in ethanol
and mmRNA(5) diluted in acidic acetate buffer (pH 4)
construct LNPs encapsulated with mmRNA. Upon mixing in acidic condition, electrostatic interactions drive the formation of inverted micelles
containing mmRNA molecules surrounded predominantly by ionizable lipid, which are then coated with PEG-lipids to form the LNPs. After LNPs formation,
the pH is raised to physiological pH, leading to LNPs with a biocompatible neutral charge. Subsequently, upon endocytosis to the acidic endosome, the
amino group of the DLin-MC3-DMA becomes positively charged, which allows the release of mmRNA molecules to the cytosol. b Representative transmission
electron microscopy (TEM) images of LNP encapsulating fLuc mmRNA (left) and IL10 mmRNA (right). c Encapsulation efficiency as measured using RiboGreen
assay. Free mmRNA concentration in Triton-permeabilized LNPs solution divided by free mmRNA concentration in intact LNPs solution. So 6-FAM attaches
to FAM-CAGGTGGAACCTCATCAGGAGATGC-BBQ and identifies it as 2019-nCoV. www.nature.com...
However, this engineered virus may not purposely replicate enough helper lipid DSPC (1), PEG-lipid (2), ionizable amino lipid DLin-MC3-DMA (3) and
cholesterol (4) for the 6-FAM to bind too. Until the viral load, meets the level for enough 6-FAM to bind too, it’s too late.
Perhaps a test for the targeting moiety (the specific antigen and Compliment that attaches to the epithelium of the alveoli), would reveal faster
As always, please refer to my signature below.
edit on V092020Tuesdaypm29America/ChicagoTue, 04 Feb 2020 21:09:23 -06001 by Violater1 because: color