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
13
Interpretation of Results and Reporting
Review patient results for unusual patterns, trends or distributions in patient results, such as an unusually high percentage
of abnormal results, or unusually high percentage of non-reactive, or indeterminate, or reactive results. Computer aided
tools should be used when available. Refer to SOP Quality Control Program and Molecular Infectious Diseases Department.
Real-Time Group Results Review and Release Process.
Report atypical or unexpected results or trends for this test to appropriate supervisory personnel, prior to releasing results.
When all controls exhibit the expected performance (Acceptance Criteria for Controls), a specimen is considered
negative if all SARS-CoV-2 markers (N1, N3) cycle threshold amplification curves do not cross the threshold and the
IPC amplification curve does cross the threshold line within the acceptance range.
When all controls exhibit the expected performance, a specimen is considered Detected for SARS-CoV-2 if all markers
(N1, N3) cycle threshold amplification curves cross the threshold line (<40.00 Ct). The IPC may or may not be positive
as described above, but the SARS-CoV-2 result is still valid.
When all controls exhibit the expected performance and the amplification curves for the SARS-CoV-2 markers (N1, N3)
and the IPC amplification curve does not cross the threshold line within the acceptance range, possible PCR inhibition
has occurred for the specimen. Specimen should be re-tested. If upon repeat testing the same situation occurs the patient
result is reported as “Indeterminate due to inhibition” (TNP1146).
When all controls exhibit the expected performance and the cycle threshold amplification curve for any one or two
markers, (N1, N3) but not all two crosses the threshold line (< 40.00 Ct), the result is inconclusive for SARS-CoV-2. The
sample should be rerun. If upon repeat testing the same situation occurs the patient result is reported as “Inconclusive”.
Specimen Result Interpretation*
(for specimens that are not pooled)
nCoV-N1
nCoV-N3
IPC
Interpretation
Actions
ND
ND
Within +/- 3 Ct of
Negative Control
NOT DETECTED
Report to public health authorities.
DET
DET
Not Applicable (+/-)
DETECTED
Report to public health authorities. Store
Samples at -70°C or colder
to refer to the
appropriate Public Health laboratory if
requested.
Only one of two SARS-
nCoV-
2 targets are
Detected
Not Applicable (+/-)
INCONCLUSIVE
Repeat extraction and RT-PCR. If the
repeated result remains Inconclusive,
Store Samples at -70°C or colder
to refer
to the appropriate Public Health
laboratory if requested.
ND
ND
Undetermined or
IPC out of range
(>3Ct)
INVALID
Repeat extraction and RT-PCR. If upon
repeat testing the same situation occurs
the patient result is reported as “Unable
to report” due to inhibition (TNP1146).