BREAKING NEWS: THE SUMMARY AEUA FOR THIS PCR TEST, SAYS THE TEST IS "USELESS" *
I located the Summary findings AEUA for a PCR test. It should be front page news because it discloses that the PCR test is ‘useless’ at providing any useful information. But getting it approved under this Emergency Use Authorization is ok, because the standard is even lower for the PCR test: it is an Accelerated Emergency Use Authorization.
Note: There is EU funding associated with some of the testing that got this approved - so there is even a conflict in this AEUA.
IN THE accelerated EUA FOR THIS PCR TEST IT ACTUALLY INDICATES THE TEST IS USELESS;
Within the Accelerated Emergency Use Authorization it says:
“Positive results do not rule out bacterial infection or co-infection with other viruses.The agent detected may not be the definite cause of disease.”
I am stunned. It is not a test for one thing. It is a test for anything. We are not talking about cycles here. We are talking about the legal documents or basis that set up the PCR test for approval. We are fundamentally attacking the Pandemic’s ‘identification’ system.
We are also educating ourselves on the different standards of testing.
I’ve heard of an Emergency Use Authorization. What is an ‘Accelerated Emergency Use Authorization?’ It turns out the AEUA has the lowest most shoddy standard for approval.
I am literally shaking my head as I draft this.
‘Does the FDA have different standards for EUA and full BLA approval?
Could you explain the difference if there is one?
Summary:
Yes, the FDA does have different standards. For this discussion, there are three standards. 2
1. A higher “established as safe and effective” standard applied to a conventional BLA.
2. A lower “believes may be effective” standard applied to an EUA.
3. A lower “reasonably likely to predict” standard applied to an “accelerated approval” BLA used to approve seasonal influenza and updated COVID-19 vaccines and which relies on surrogate endpoints whose establishment does not require the “substantial evidence” standard.”
I pulled this information from THIS Congressional document.
So basically this PCR test was approved on the basis of this lowest standard of ‘reasonably likely to predict’ standard. And even within that standard they say the positive result for the PCR test doesn’t rule out other viruses, bacterial infection and whatever is detected may not be the cause of disease. I call that *Useless . It is likely to predict- no qualify that- it is reasonably likely to predict…. what? a bunch of things.
THAT IS NOT A TEST FRIENDS.
I think we just broke the PCR TEST.
“Diatherix Eurofins SARS-CoV-2 Assay EUA Summary – Updated September 22, 2020
ACCELERATED EMERGENCY USE AUTHORIZATION (EUA) SUMMARY SARS-CoV2 RT-PCR Assay
(Diatherix Eurofins Laboratory) For In vitro Diagnostic Use Rx Only
For use under Emergency Use Authorization (EUA) only (The SARS-CoV-2 RT-PCR assay will be performed at the Diatherix Eurofins Laboratory, certified under the Clinical Laboratory Improvement Amendments of 1988 (CLIA), 42 U.S.C. §263a as per Laboratory Instructions for Use that was reviewed by the FDA under this EUA.)
INTENDED USE
The SARS-CoV-2 assay is a PCR test intended for the qualitative detection of nucleic acid from SARS-CoV-2 in nasal swabs, nasopharyngeal swabs/wash, oropharyngeal swabs/throat swabs, bronchial aspirates, or sputum from individuals in which identification of SARS-CoV-2 would provide relevant clinical information to the individual and to the healthcare provider. Testing is limited to Diatherix Laboratories, LLC, a College of American Pathologists (CAP)-accredited, and Clinical Laboratory Improvement Amendments of 1988 (CLIA), 42 U.S.C. §263a certified high-complexity laboratory.
Results are for the detection and identification of SARS-CoV-2 RNA. SARS-CoV-2 RNA is generally detectable in respiratory specimens during the acute phase of infection.
Positive results are indicative of the presence of SARS-CoV-2 RNA. Clinical correlation with patient history and other diagnostic information is necessary to determine patient infection status.
Positive results do not rule out bacterial infection or co-infection with other viruses. The agent detected may not be the definite cause of disease.
Laboratories within the United States and its territories are required to report all positive results to the appropriate public health authorities.
Negative results do not preclude SARS-CoV-2 infection and should not be used as the sole basis for patient management decisions.
Negative results must be combined with clinical observations, patient history, and epidemiological information. The assay is intended for use under the Food and Drug Administration’s Emergency Use Authorization.
Diatherix Eurofins SARS-CoV-2 Assay EUA Summary – Updated September 22, 2020
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DEVICE DESCRIPTION AND TEST PRINCIPLE
Diatherix’s Target-Enriched Polymerase Chain Reaction is a nested, end-point PCR technology that allows SARS-CoV-2 detection through target enrichment and amplification.
The reaction process includes enrichment and tagging of the target, followed by traditional PCR amplification. First, nucleic acid is extracted from the sample. Each targeted pathogen is then amplified by low concentration nested gene-specific primers that are designed to enrich the targets during the initial PCR cycles. Inside nested primers have a unique tag sequence complementary to proprietary SuperPrimers (Fs and Rs), which are included in the primer mix. The universal SuperPrimers are used to amplify all targets by annealing to the complementary tag sequence on the inside nested primers.
The reverse primer of the SuperPrimer set is labeled with biotin, which is incorporated into the resultant PCR product. The schematic below describes this procedure:
Figure 1. Reaction Process. Low concentration nested gene-specific primers are (Fo – forward out; Fi – forward in; Ri– reverse in; and Ro – reverse out) designed to enrich the targets during the initial PCR cycles. Later in the procedure, a pair of universal SuperPrimers (Fs and Rs) is used to amplify all targets. The Rs primer is labeled with biotin for subsequent detection.
Following PCR, amplicons are hybridized onto a Diatherix microarray in which a detection probe specific to the assay target is attached. Hybridization is followed by incubation with streptavidin-phycoerythrin (SA-PE), which binds to the biotinylated amplicon and emits fluorescence upon excitation. Results for SARS-CoV-2 are reported as ‘Detected’ or ‘Not Detected’ based on fluorescence intensity above background.
INSTRUMENTS USED WITH TEST
The Diatherix SARS-CoV-2 assay has been validated to be used with the KingFisher Flex System (Thermo Fisher Scientific, Waltham, MA) utilizing extraction reagents from either Qiagen (ClearMag) or Omega Bio-tek (MagBind Viral DNA/RNA). RT-PCR reactions have been validated on and can be performed on the Applied Biosystems (ABI) GeneAmp 9700 or the Veriti 96-well Thermal Cycler (Thermo Fisher Scientific). Postamplification hybridization washes are performed using a Wellwash Versa Microplate Washer (Thermo Fisher Scientific). Fluorescence signals are analyzed on the SensoSpot Fluorescence Low Density Microarray Analyzer (Sensovation AG, Radolfzell, Germany).
Diatherix Eurofins SARS-CoV-2 Assay EUA Summary – Updated September 22, 2020
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REAGENTS AND MATERIALS
Table 1. Preferred vendors and catalog numbers
(LL: SEE DOWNLOADED DOCUMENT)
Diatherix Eurofins SARS-CoV-2 Assay EUA Summary – Updated September 22, 2020
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CONTROLS TO BE USED WITH THE SARS-CoV-2 ASSAY
For SARS-CoV-2 patient sample testing, the following controls are required: Negative Extraction control consisting of H1N1-09 organism, Positive Control consisting of SARS-CoV-2 IVT RNA, and an Internal Amplification Control. A PCR negative control must also be run with every batch.
A description of each control is below:
SARS-CoV-2 Positive Control:
One positive control is run on each plate. This control is designed to assess the integrity of the PCR run. The positive control consists of SARS-CoV-2 in vitro transcribed RNA from a partial sequence of the S gene diluted in Simulated Lung Fluid (SLF, also known as Gamble’s Solution)1 for a final concentration of 100 copies/µL.
Internal Control (IC):
An internal control is run with every patient sample from extraction. This control is a synthetic oligonucleotide sequence that is complementary to universal primers included in the assay. The control is spiked into the elution buffer used during nucleic acid extraction and is intermingled with the nucleic acids of patient samples during the final step of nucleic acid extraction. The IC is carried throughout the remainder of the assay steps.
NTC (No Template Control):
This control is run in every batch (up to 96 samples) to rule out contamination of reagents with target nucleic acid. The no template control is a PCR negative control and consists of 4 µL of the nuclease-free water provided in the Luna Universal One-Step RT-qPCR enzyme kit.
SARS-CoV-2 Negative Extraction Control:
The SARS-CoV-2 negative extraction control is run on each plate. This control consists of the H1N1-09 organism. This is a process control that monitors for the entire lysis, extraction, amplification, and detection reactions.
INTERPRETATION OF RESULTS
1) SARS-CoV-2 RT-PCR test Controls – Positive, Negative, and Internal:
Positive Control: Amplification must yield a positive signal (above cutoff) for the SARS-CoV-2 target (NCOVDe2 Detection sequence) and yield negative signal for all other targets. If the extraction control fails, the cause of the failure will be investigated and the run repeated, if warranted.
1 Marques, M., Loebenberg, R., and Almukainzi, M. 2011. Simulated Biological Fluids with Possible Application in Dissolution Testing. Dissolution Technologies.15-28. <dx.doi.org/10.14227/DT180311P15> Diatherix Eurofins SARS-CoV-2 Assay EUA Summary – Updated September 22, 2020
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Internal Control: The internal control must be detected for a negative result to be reported. For specimens in which the IC is not detected, it will be assumed that the sample includes a PCR inhibitor, and the sample will be repeated from extraction.
No Template Control: The PCR negative control must have all target signals below their respective cutoff for the run to pass. If the negative control contains positive signals for any of the targets, the run will be repeated.
SARS-CoV-2 Negative Extraction Control: This control should only be positive for the H1N1 targets and negative for all other targets. If the extraction control fails, the cause of failure will be investigated and the run repeated, if warranted.
If any batch control (NTC or Extraction Control) fails, the Laboratory Manager or Technical Supervisor will be notified. If an individual patient sample control fails, the patient report will be held and repeated. If the sample fails upon repeat testing, the affected sample will be reported as a failed test.
2) Examination and Interpretation of Patient Specimen Results:
Results for the SARS-CoV-2 target are reported as “Detected” or “Not Detected” based on signal intensity above background (cutoff), measured in Relative Fluorescence Units (RFU).
1. Positive Specimens:
Specimens with a SARS-CoV-2 signal intensity above the pre-defined cutoff will be considered detected.
2. Negative Specimens:
Specimens with a SARS-CoV-2 signal intensity level below the pre-defined cutoff will be considered not detected only if the internal control is also detected.
3. Failed Test:
If a signal intensity cannot be generated for a well, the patient sample is considered a “hybridization failure” and the run must be repeated from hybridization. If the sample fails upon retesting, the affected sample will be reported as a failed test.
Table 2. Interpretation of Patient Results.
Target I (LL see downloaded document)
PERFORMANCE EVALUATION
1) Analytical Sensitivity:
Limit of Detection (LoD):
The analytical sensitivity of the SARS-CoV-2 assay was determined in Limit of Detection (LoD95) studies. LoD95 is defined as the concentration that can accurately be detected 95% of the time. Since no quantified viral isolates were available at the time of the study, assays were tested from extraction with characterized genomic RNA (Human 2019-nCoV strain 2019-nCoV/Italy-INMI12 ) and stocks of in vitro transcribed (IVT) RNA for a partial sequence of the spike (S) gene. Genomic RNA and IVT RNA of known titers (copies/µL) were spiked into non-reactive, pooled sputum clinical matrix and simulated lung fluid (SLF) to mimic respiratory swab specimens. Nucleic acid extraction was performed on the KingFisher Flex extraction platform.
To first determine performance of the assay, 10-fold serial dilutions of IVT RNA in SLF were prepared (ranging from 1010 copies/µL to 100 copies/µL) and extracted in triplicate using Qiagen ClearMag extraction reagents. PCR was performed on the Veriti 96 well thermocycler.
Once assay performance was assessed, the same test conditions were used to determine broad-range LoD for the assay in clinical matrix. Broad range LoD was determined by testing triplicate samples of IVT RNA and genomic RNA spiked in pooled sputum from 103 to 100 copies/µL on the ABI 9700 thermocycler.
The LoD was confirmed to be 1 copy/µL based on a positivity rate of 95% upon testing 60 replicates in pooled sputum matrix (57/60). Samples for the LoD confirmation study were extracted using the Qiagen ClearMag or Omega Bio-tek kits.
2) Analytical Inclusivity
An alignment was performed with the oligonucleotide primer and detection sequences designed for the Diatherix SARS-CoV-2 assay using all publicly available sequences for SARS-CoV-2 found in NCBI Virus Resource and GenBank as of March 15, 2020 to predict the inclusivity of the target. All alignments show 100% identity to the primer sequences utilized by the SARS-CoV-2 assay.
3) Cross-Reactivity
A total of 31 viral strains, 34 bacterial strains, and two fungal strains were wet tested for cross-reactivity against the SARS-CoV-2 assay (Table 3). The organisms were
2 This work was supported by the European Virus Archive Goes Global (EVAg) project that has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 653316.
Diatherix Eurofins SARS-CoV-2 Assay EUA Summary – Updated September 22, 2020
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selected for their likelihood to be found in the respiratory tract, for genetic similarities, or for likelihood of carriage within the clinical matrix. For this testing, titered organisms were prepared in mixes containing up to six microorganisms at the concentrations listed below, spiked into SLF and extracted utilizing Qiagen ClearMag reagents in at least triplicate. RT-PCR reactions were performed on the Veriti 96-well thermocycler. No false positive results indicating cross-reactivity were observed.”
Read the FULL ACCELERATED EMERGENCY USE AUTHORIZATION HERE:
This should break the hegemony of the PCR test and shatter it. Please share like comment and move this substack. Thank-you for your support.
The gentleman who developed the PCR test (Carey) and won a Nobel Prize for it was very clear in an interview some years ago that the PCR test was not useful for this kind of testing and the higher the cycles the less reliable it became (my memory may be sketchy on the finer points of this issue), and he was also NO FAN of Anthony Fauci…amazingly, he up and died the fall of 2019, just before the pandemic struck! How convenient for the Event 201 coordinators…
At best, the PCR could only detect if one had been exposed to the virus but exposure does not mean infection and infection does not mean sick. Apart from covid, a single positive test result is never enough to diagnose anything. Not in the human body or plant operations.