For in vitro diagnostic use. Others cobas MPXV IVD cobas® MPXV For use under Emergency Use Authorization (EUA) only PID00000469 Qualitative assay for use on the cobas® 6800/8800 Systems 09863338190 KIT COBAS 68/8800 MPXV 192T KIT COBAS 68/8800 MPXV 00875197007176 Reagents, kits 1 kit 192 tests true cobas® MPXV for use on the cobas® 6800/8800 Systems (cobas® MPXV) is a real-time PCR assay for the qualitative detection of DNA from Monkeypox virus (MPXV, clade I/II) in human lesion swab specimens (i.e., swabs of acute pustular or vesicular rash) from individuals suspected of monkeypox infection by their healthcare provider. Testing is limited to laboratories certified under the Clinical Laboratory Improvement Amendments of 1988 (CLIA), 42.U.S.C. §263a, that meet requirements to perform moderate or high complexity tests.Results are for the identification of monkeypox virus (clade I/II) DNA, which is generally detectable in human pustular or vesicular lesion specimens during the acute phase of infection. Positive results are indicative of the presence of monkeypox virus (clade I/II) DNA; 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. Negative results obtained with this device do not preclude monkeypox virus (clade I/II) infection and should not be used as the sole basis for treatment or other patient management decisions. Negative results must be combined with clinical observations, patient history, and epidemiological information.Laboratories within the United States and its territories are required to report test results to the appropriate public health authorities. cobas® MPXV is intended for use by qualified clinical laboratory personnel specifically instructed and trained in the techniques of real-time PCR and on the use of the cobas® 6800/8800 Systems.cobas® MPXV is only for use under the Food and Drug Administration’s Emergency Use Authorization. en cobas® MPXV is based on fully automated sample preparation (nucleic acid extraction and purification) followed by PCR amplification and detection. The cobas® 6800/8800 Systems consist of the sample supply module, the transfer module, the processing module, and the analytic module. Automated data management is performed by the cobas® 6800/8800 System software, which assigns test results for all tests. Results can be reviewed directly on the system screen, and printed as a report.Nucleic acid from patient samples and added Internal Control DNA (DNA IC) molecules are simultaneously extracted. Nucleic acid is released by addition of proteinase and lysis reagent to the sample. The released nucleic acid binds to the silica surface of the added magnetic glass particles. Unbound substances and impurit ies, such as denatured protein, cellular debris and potential PCR inhibitors, are removed with subsequent wash steps and purified nucleic acid is eluted from the magnetic glass particles with elution buffer at elevated temperature. External controls (positive and negative) are processed in the same way.Selective amplification of target nucleic acid from the sample is achieved by the use of three sets of forward and reverse primers targeting the MPXV F3L gene, the MPXV B21R/B22R gene, and the human β-globin gene, which were selected to hybridize to highly conserved regions of the genomic nucleic acid. Selective amplification of DNA Internal Control is achieved by the use of non-competitive sequence specific forward and reverse primers which have no homology with the Monkeypox virus or human genomes. A thermostable DNA polymerase enzyme is used for both reverse- transcription and amplification.The cobas® MPXV master mix contains detection probes which are specific for MPXV (targeting MPXV genes F3L and B21R/B22R, labeled with the same fluorescent dye), the human β-globin gene and Internal Control nucleic acid. The MPXV, β-globin and Internal Control detection probes are each labeled with unique fluorescent dyes that act as a reporter. Each probe also has a second dye which acts as a quencher. When not bound to the target sequence, the fluorescent signals of the intact probes are suppressed by the quencher dye. During the PCR amplification step, hybridization of the probes to the specific single-stranded DNA template results in cleavage of the probe by the 5' to 3' exonuclease activity of the DNA polymerase resulting in separation of the reporter and quencher dyes and the generation of a fluorescent signal. With each PCR cycle, increasing amounts of cleaved probes are generated and the cumulative signal of the reporter dye increases concomitantly. Each reporter dye is measured at defined wavelengths, which enables simultaneous detection and discrimination of the amplified MPXV targets, β-globin and the DNA Internal Control. The master mix includes deoxyuridine triphosphate (dUTP), instead of deoxythimidine triphosphate (dTTP), which is incorporated into the newly synthesized DNA (amplicon). Any contaminating amplicons from previous PCR runs are destroyed by the AmpErase enzyme [uracil-N-glycosylase], which is included in the PCR mix, when heated in the first thermal cycling step. However, newly formed amplicons are not destroyed since the AmpErase enzyme is inactivated once exposed to temperatures above 55°C. en