Presentation on theme: "and Respiratory Viruses in Multiple Specimen Types"— Presentation transcript:
1 Evaluation of the Film Array Respiratory Panel for Detection of Mycoplasma pneumoniae and Respiratory Viruses in Multiple Specimen TypesEvaluation of the Film Array Respiratory Panel for Detection of Mycoplasma pneumoniaeand Respiratory Viruses in Multiple Specimen TypesTitle HereChristine Robinson1, Kristin Pretty1, Daniel Olson2, and Samuel Dominguez2Departments of Pathology and Laboratory Medicine1 and Pediatrics2 , Children’s Hospital Colorado1,2 and University of Colorado2,Anschutz Medical Campus, COChristine Robinson1, Kristin Pretty1, Daniel Olson2, and Samuel Dominguez2Departments of Pathology and Laboratory Medicine1 and Pediatrics2 , Children’s Hospital Colorado1,2 and University of Colorado2,Anschutz Medical Campus, COBACKGROUNDRESULTSRESULTSFILM ARRAY RESP. PANEL - TESTING OF MULTIPLE SPECIMEN TYPES FOR VIRUSFILM ARRAY RESP. PANEL - TESTING OF MULTIPLE SPECIMEN TYPES FOR MPORGANISM AND SIGNIFCANCEMycoplasma pneumoniae (Mp) is a bacterial cause of community-acquired pneumonia and occasional extra-pulmonary disease in school-age children and young adults. Most cases are mild but some are hospitalized. Stevens Johnson Syndrome (SJS), a rare but serious disorder involving skin and mucous membranes usually triggered by medication, can also be induced by Mp. Circulation usually is endemic, but outbreaks can occur. Throat swabs (TS) are the usual specimen sent to detect Mp by PCR.A. Many Specimen Types TestableB. Validation of FA-RP vs. Luminex RVPA. Mp Detected in Many Specimen TypesB. Validation of FA-RP Mp PCR vs. Other PCRs20 NPS45 NPW40 TA40 BALLuminex RVP*Luminex RVP**PosNegTotalFilm Array RP24125286342045445564064158048029646675600DiscrepancyMissed 1 AdV Detected 1 add'l RVFA missed 1 AdVNoneSensitivity96%98.70%100%Specificity99.80%99.10%Agreement99.50%98.90%Film Array RPSpecimen TypesMp PosTestedPos RateFDA ClearedNPS97411.2%Lab-ValidatedNW6434211.9%BAL104412.3%TA22220.1%Other1040.0%Total76 *41881.8%Total Tested85 *49291.7%* 89% of Mp FA positives were in lab-validated specimensConfirmatory Mp PCR ( CDC and/or Focus)Same Specimen as FADifferent Specimen as FAPosNegTotalFilm Array RP79483162181188512220187103Sensitivity100%100.00%Specificity96.70%97.70%Agreement98.00%98%M. pneumoniaeScanning electron micrograph.Hospitalized child with severe SJS skin and eye involvement* IVD version **RUO versionOUTBREAKAn outbreak of Mp-associated respiratory disease and unusual cluster of Mp- associated SJS was identified in Denver-area children in late 2013, prompting an on-site investigation by CDC. Most of these infections were detected by the Film Array Respiratory Panel (FA RP, Biofire Diagnostics), an FDA-cleared PCR for 17 respiratory viruses and 3 bacteria in nasopharyngeal swabs (NPS). Specimens other than NPS were Mp-positive, although we had not yet validated reporting of bacteria from this test. Other cases were detected by Mp PCR of TS at Focus Diagnostics Laboratory.MYCOPLASMA PNEUMONIAEFilm Array RP Detects OutbreakUse of FA for respiratory virus detection begins in early 2013; bacteria to be validated later. If Mp detected, specimen sent to Focus for confirmation before reporting. Mp detection increases significantly in autumn.C. Agreement between CDC and Focus Mp PCRs88% agreement when the same 50 FA-positive specimens tested by both laboratories83% positive agreement when different FA-positive specimens from the same patient testedCDC PCR identified 4 Mp-positive specimens resulted as Mp negative at Focus; Focus detected no additional positives .Organisms in FA Respiratory PanelViruses BacteriaInfluenza A, H1, H1 2009, H3 Mycoplasma pneumoniaeInfluenza B Chlamydophila pneumoniaeRSV Bordetella pertussisHuman metapneumovirusAdenovirusParainfluenza 1-4Coronaviruses 229E, OC43, HKU1, NL63Rhinovirus/enterovirusAll FA Results; Feb 2013-Feb 2014Film Array SystemOBJECTIVESDescribe a clinical validation of FA for detection of respiratory viruses and Mp in multiple specimen types including NPS, nasopharyngeal washes (NPW), tracheal aspirates (TA), and bronchoalveolar lavage (BAL), using specimens collected during a Mp outbreak.Association of MP with SJSLink between SJS and Mp observed. CDC investigates on-site; verifies community-wide Mp outbreak and association of Mp with some SJS cases.CONCLUSIONSMETHODSRESPIRATORY VIRUS VALIDATION: Banked or fresh NPW, TA, and BAL positive by Luminex Respiratory Virus Panel (LUM, IUO version), donated LUM-positive NPS, and negative specimens spiked with virus were combined into 2-specimen pools and tested by FA v1.6. The number of expected and observed analytes for each pool was scored.Mp – CLINICAL VALIDATION: 288 banked or fresh respiratory specimens positive or negative for Mp by FA were sent to the CDC’s Mycoplasma Laboratory only (19 pos), Focus Reference Laboratory (16 pos, 203 neg), or both laboratories (50 pos) for Mp PCR. Mp strain typing by MLVA and resistance testing was performed at CDC.Mp – ANALYTICAL VALIDATION: In progressSpecimens other than NPS can be tested for viruses by Film Array RP and resulted with confidenceDetection of bacteria compared to viruses in respiratory tract specimens of children is lowAn outbreak of Mp tract disease identified by Mp PCR of respiratory tract specimens occurred in late 2013 in Denver area children. Some cases were associated with Stevens-Johnson DiseaseClinical detection of M. pneumoniae by Film Array RP was validated in multiple specimen types7 Mp-associated probable SJS cases identified Aug-Nov 2013; 5 considered confirmed. All had severe disease; several had recurrent disease.3 different strains of Mp were circulating; no single strain was identified in SJS patientsAll 4 Mp from SJS patients were macrolide sensitive; 7% of 45 Mp from non-SJS patients and contacts were resistant.