Instruction Manual
Table Of Contents
- Intended Use
- Summary and Explanation
- Principles of the Procedure
- Materials Required (Provided)
- DTT solution 500mM
- SARS-CoV-2 PCR Mix
- Equipment and Supplies
- Warnings and Precautions
- Specimen Collection, Handling, and Storage
- Procedure
- For viscous samples (sputum and bronchial wash): Performed inside BSC 2
- NOTE: In the event that the test system becomes inoperable, notify supervision or designee for further direction. Patient specimens must be stored in a manner that maintains the integrity of the specimen.
- Quality Control
- Limitations
- Conditions of Authorization for the Laboratory
- Performance Characteristics
- References
15
Human coronavirus HKU1
HKU1
Human coronavirus NL63
NL63
Adenovirus (e.g. C1 Ad. 71)
Type 3
Human Metapneumovirus
8, Peru6-2003
Parainfluenza virus 1-4
Parainfluenza 1-4
Influenza A
A/Brisbane/10/07
Influenza B
B/Florida/02/06
Respiratory syncytial virus
A
Rhinovirus
1A
Chlamydia pneumoniae
M129
Bordetella pertussis
A639
Mycoplasma pneumoniae
M129
4) Interfering substances study
The assay uses conventional well-established nucleic acid extraction methods and based on our experience with
other similar assays, e.g. Influenza A and B Real-Time PCR. We do not anticipate interference from common
endogenous substances. Interference studies have not been performed for this assay.
5) In silico cross-reactivity testing
Cross-reactivity is defined as the amplification and detection of related viruses or other pathogens by the SARS-
CoV-2 RNA Qualitative RT-PCR assay. CDC determined that the 2019-nCoV rRT-PCR assay N1 and N3, designed
for the detection of SARS-CoV-2, showed no significant combined homologies with human genome, other
coronaviruses (with the exception of N3 with SARS homology), or human microflora that would predict potential
false positive rRT-PCR results. The N3 RT-PCR is expected to cross-react with human SARS coronavirus and bat
SARS-like coronaviruses. Quest Diagnostics is using the same sequences as CDC, therefore, additional in silico
studies were determined to be unnecessary.
6) Specimen Stability and Fresh-frozen Testing
Quest Diagnostics intends to follow the CDC’s specimen collection and transport guidance contained in CDC EUA
IFU under Specimen Collection, Handling, and Storage and the CDC website for guidance on specimen collection
handling and storage (
https://www.cdc.gov/coronavirus/2019-nCoV/lab/guidelines-clinical-specimens.html
).
7) Inter-assay Precision
Inter-assay precision is defined as the reproducibility of a sample between assay runs and was evaluated by testing
3 replicates of 3 separate precision standards in three separate runs using the SARS-CoV-2 RNA Qualitative RT-
PCR assay. The precision standards were aliquots of the same standards prepared as described in the Intra-assay
Precision section. Precision standard replicate CV’s ranged from 0.1% to 0.7%, with mean overall inter-assay
precision equal to 0.3%. The individual precision standard replicate results were within 0.20 detection cycles of their
respective mean values.