Clyde

No problem. The HERV-K102 ddCt quantitative PCR (qPCR) method that I actually developed while in the Blood Safety Program was for the screening of donor blood, for cases involving transfusion, for cases of transplantation, and especially suitable for cases of detecting xenozoonosis (ie., emerging pathogens) in xenotransplantation It is based on the notion that HERV-K102 particle production may be an early indicator of: an infection, a tumor/cancer, foreign DNA/RNA and/or a toxin. Any of the above might actually be increased in persons who received the spike mRNA gene therapy LNPs and does not necessarily need to be detectable in the blood per se ie., spike protein. The main principle of the method is to compare the ratio of the HERV-K102 pol signal to the 18S RNA signal (the control for genomic equivalents) of isolated DNA (the QIAmp UltraSens Virus particle Isolation kit that also copurifies genomic DNA from plasma). The method utilizes a reference genome (male genomic DNA of a known concentation) to enable quantification. Here is the reference. Laderoute MP, Giulivi A, Larocque L, et al. The replicative activity of human endogenous retrovirus K102 (HERV-K102) with HIV viremia. AIDS. 2007 Nov 30;21(18):2417-24. Generally when there is an active infection the ddCt often indicates about 10 (12) HERV-K102 particles per ml of plasma or a ddCt of 10 (9). When performed on 30 normal healthy controls validated not to have any HERV-K102 particles and no antibody to HERV-K102 envelope, the ddCt was 0.88+/- 0.37. The induction of HERV-K102 particles can be very quick such as zero particles per ml of plasma to 2.55 X 10(11) cDNA containing particles in 84 hours. My method uses an internal sequence to validate the proper sequence has been amplified (after 35 cycles), and to provide real time assessment during each cycle. My method seems to be very accurate as discussed elsewhere: Laderoute MP. Clues to finding correlates of risk/protection for HIV-1 vaccines [version 2; peer review: 2 approved with reservations] F1000 Research. 2018, 6:868. doi.org/10.12688/f1000…. If the test is performed with Amperase-UNG(TM) in the buffer, this dissolves the cDNA in HERV-K102 particles (it is a foamy retrovirus and foamy non-pathogenic retroviruses have cDNA genomes due to their opposite lifecycle to pathogenic retroviruses). Recall that cDNA contains uracil not thymine. So when 2 qPCRS are done, one with the Amp-UNG and the other not, this allows one to be confident that one has indeed measured the cDNA of HERV-K102 particles (because the cDNA signal of the HERV-K102 in the particles disappeasrs in the presence of Amp-UNG) . This method can also be used to quantitate integration levels of HERV-K102 which may indicate a recent infection/toxin. For example, a special cohort of women known to be resistant to HIV-1 acquisition for at least the past 3 years, scored positive for elevated integration (average was 5 fold over normal) which was not found in HIV-1 patients. Laderoute MP, Larocque LJ, Giulivi A, Diaz-Mitoma F. Further evidence that human endogenous retrovirus K102 is a replication competent foamy virus that may antagonize HIV-1 replication. Open AIDS J. 2015 Dec 7;9:112-22. doi: 10.2174/1874613601509010112. Others have found that increased integration of HERV-K102 in lung adenocarcinomas relates to higher survival of the host (all-cause mortality). [Ng KW, Boumelha J, Enfield KSS, et al. Antibodies against endogenous retroviruses promote lung cancer immunotherapy. Nature. 2023 Apr;616(7957):563-573. doi: 10.1038/s41586-023-05771-9. ]. Preliminary evidence suggests even small exposures to pesticides might be detected with this method in the range of 2.5 to say 5.0. The beta testing was never done despite the patents as the Director (a Hematologist) left PHAC suddenly and the new Director had no interest in blood safety and could not see the significance. So clearly the testing needs to be properly evaluated for approval by the FDA.