Validation of a novel molecular diagnostic panel for pediatric musculoskeletal infections: Integration of the Cepheid Xpert MRSA/SA SSTI and laboratory-developed real-time PCR assays for clindamycin resistance genes and Kingella kingae detection

Pathogen detection in pediatric patients with musculoskeletal infections relies on conventional bacterial culture, which is slow and can delay antimicrobial optimization. The ability to rapidly identify causative agents and antimicrobial resistance genes in these infections may improve clinical care...

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Veröffentlicht in:	Journal of microbiological methods Jg. 156; S. 60 - 67
Hauptverfasser:	Searns, Justin B., Robinson, Christine C., Wei, Qi, Yuan, Ji, Hamilton, Stacey, Pretty, Kristin, Donaldson, Nathan, Parker, Sarah K., Dominguez, Samuel R.
Format:	Journal Article
Sprache:	Englisch
Veröffentlicht:	Netherlands Elsevier B.V 01.01.2019
Schlagworte:	Anti-Bacterial Agents - therapeutic use antibiotic resistance antibiotic resistance genes Bacterial Proteins - genetics Child children clindamycin Clindamycin - therapeutic use Colorado Diagnostics Drug Resistance, Bacterial Female hospitals Humans Infection Kingella kingae Kingella kingae - genetics Kingella kingae - isolation & purification Male methicillin-resistant Staphylococcus aureus Methyltransferases - genetics microbial detection musculoskeletal system Neisseriaceae Infections - diagnosis Osteoarthritis - microbiology Osteomyelitis Osteomyelitis - microbiology patients Pediatric phenotype quantitative polymerase chain reaction Septic arthritis Colorado Osteomyelitis Infection Diagnostics NPV Septic arthritis PPV MDP qPCR Pediatric CHCO
ISSN:	0167-7012, 1872-8359, 1872-8359
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Abstract	Pathogen detection in pediatric patients with musculoskeletal infections relies on conventional bacterial culture, which is slow and can delay antimicrobial optimization. The ability to rapidly identify causative agents and antimicrobial resistance genes in these infections may improve clinical care. Convenience specimens from bone and joint samples submitted for culture to Children's Hospital Colorado (CHCO) from June 2012 to October 2016 were evaluated using a “Musculoskeletal Diagnostic Panel” (MDP) consisting of the Xpert MRSA/SA SSTI real-time PCR (qPCR, Cepheid) and laboratory-developed qPCRs for Kingella kingae detection and erm genes A, B, and C which confer clindamycin resistance. Results from the MDP were compared to culture and antimicrobial susceptibility testing (AST) results. A total of 184 source specimens from 125 patients were tested. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the Xpert MRSA/SA SSTI compared to culture and AST results were 85%, 98%, 93%, and 95% respectively for MSSA and 82%, 100%, 100%, and 99% for MRSA. Compared to phenotypic clindamycin resistance in S. aureus isolates, the erm A, B, and C gene PCRs collectively demonstrated a sensitivity, specificity, PPV, and NPV of 80%, 96%, 67%, and 98%. In comparison to clinical truth, Kingella PCR had a sensitivity, specificity, PPV, and NPV of 100%, 99.5%, 100%, and 100%. This novel MDP offers a rapid, sensitive, and specific option for pathogen detection in pediatric patients with musculoskeletal infections.
AbstractList	Pathogen detection in pediatric patients with musculoskeletal infections relies on conventional bacterial culture, which is slow and can delay antimicrobial optimization. The ability to rapidly identify causative agents and antimicrobial resistance genes in these infections may improve clinical care.Convenience specimens from bone and joint samples submitted for culture to Children's Hospital Colorado (CHCO) from June 2012 to October 2016 were evaluated using a “Musculoskeletal Diagnostic Panel” (MDP) consisting of the Xpert MRSA/SA SSTI real-time PCR (qPCR, Cepheid) and laboratory-developed qPCRs for Kingella kingae detection and erm genes A, B, and C which confer clindamycin resistance. Results from the MDP were compared to culture and antimicrobial susceptibility testing (AST) results.A total of 184 source specimens from 125 patients were tested. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the Xpert MRSA/SA SSTI compared to culture and AST results were 85%, 98%, 93%, and 95% respectively for MSSA and 82%, 100%, 100%, and 99% for MRSA. Compared to phenotypic clindamycin resistance in S. aureus isolates, the erm A, B, and C gene PCRs collectively demonstrated a sensitivity, specificity, PPV, and NPV of 80%, 96%, 67%, and 98%. In comparison to clinical truth, Kingella PCR had a sensitivity, specificity, PPV, and NPV of 100%, 99.5%, 100%, and 100%.This novel MDP offers a rapid, sensitive, and specific option for pathogen detection in pediatric patients with musculoskeletal infections. Pathogen detection in pediatric patients with musculoskeletal infections relies on conventional bacterial culture, which is slow and can delay antimicrobial optimization. The ability to rapidly identify causative agents and antimicrobial resistance genes in these infections may improve clinical care. Convenience specimens from bone and joint samples submitted for culture to Children's Hospital Colorado (CHCO) from June 2012 to October 2016 were evaluated using a “Musculoskeletal Diagnostic Panel” (MDP) consisting of the Xpert MRSA/SA SSTI real-time PCR (qPCR, Cepheid) and laboratory-developed qPCRs for Kingella kingae detection and erm genes A, B, and C which confer clindamycin resistance. Results from the MDP were compared to culture and antimicrobial susceptibility testing (AST) results. A total of 184 source specimens from 125 patients were tested. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the Xpert MRSA/SA SSTI compared to culture and AST results were 85%, 98%, 93%, and 95% respectively for MSSA and 82%, 100%, 100%, and 99% for MRSA. Compared to phenotypic clindamycin resistance in S. aureus isolates, the erm A, B, and C gene PCRs collectively demonstrated a sensitivity, specificity, PPV, and NPV of 80%, 96%, 67%, and 98%. In comparison to clinical truth, Kingella PCR had a sensitivity, specificity, PPV, and NPV of 100%, 99.5%, 100%, and 100%. This novel MDP offers a rapid, sensitive, and specific option for pathogen detection in pediatric patients with musculoskeletal infections. Pathogen detection in pediatric patients with musculoskeletal infections relies on conventional bacterial culture, which is slow and can delay antimicrobial optimization. The ability to rapidly identify causative agents and antimicrobial resistance genes in these infections may improve clinical care.BACKGROUNDPathogen detection in pediatric patients with musculoskeletal infections relies on conventional bacterial culture, which is slow and can delay antimicrobial optimization. The ability to rapidly identify causative agents and antimicrobial resistance genes in these infections may improve clinical care.Convenience specimens from bone and joint samples submitted for culture to Children's Hospital Colorado (CHCO) from June 2012 to October 2016 were evaluated using a "Musculoskeletal Diagnostic Panel" (MDP) consisting of the Xpert MRSA/SA SSTI real-time PCR (qPCR, Cepheid) and laboratory-developed qPCRs for Kingella kingae detection and erm genes A, B, and C which confer clindamycin resistance. Results from the MDP were compared to culture and antimicrobial susceptibility testing (AST) results.METHODSConvenience specimens from bone and joint samples submitted for culture to Children's Hospital Colorado (CHCO) from June 2012 to October 2016 were evaluated using a "Musculoskeletal Diagnostic Panel" (MDP) consisting of the Xpert MRSA/SA SSTI real-time PCR (qPCR, Cepheid) and laboratory-developed qPCRs for Kingella kingae detection and erm genes A, B, and C which confer clindamycin resistance. Results from the MDP were compared to culture and antimicrobial susceptibility testing (AST) results.A total of 184 source specimens from 125 patients were tested. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the Xpert MRSA/SA SSTI compared to culture and AST results were 85%, 98%, 93%, and 95% respectively for MSSA and 82%, 100%, 100%, and 99% for MRSA. Compared to phenotypic clindamycin resistance in S. aureus isolates, the erm A, B, and C gene PCRs collectively demonstrated a sensitivity, specificity, PPV, and NPV of 80%, 96%, 67%, and 98%. In comparison to clinical truth, Kingella PCR had a sensitivity, specificity, PPV, and NPV of 100%, 99.5%, 100%, and 100%.RESULTSA total of 184 source specimens from 125 patients were tested. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the Xpert MRSA/SA SSTI compared to culture and AST results were 85%, 98%, 93%, and 95% respectively for MSSA and 82%, 100%, 100%, and 99% for MRSA. Compared to phenotypic clindamycin resistance in S. aureus isolates, the erm A, B, and C gene PCRs collectively demonstrated a sensitivity, specificity, PPV, and NPV of 80%, 96%, 67%, and 98%. In comparison to clinical truth, Kingella PCR had a sensitivity, specificity, PPV, and NPV of 100%, 99.5%, 100%, and 100%.This novel MDP offers a rapid, sensitive, and specific option for pathogen detection in pediatric patients with musculoskeletal infections.CONCLUSIONSThis novel MDP offers a rapid, sensitive, and specific option for pathogen detection in pediatric patients with musculoskeletal infections.
Author	Dominguez, Samuel R. Hamilton, Stacey Pretty, Kristin Wei, Qi Robinson, Christine C. Yuan, Ji Searns, Justin B. Parker, Sarah K. Donaldson, Nathan
Author_xml	– sequence: 1 givenname: Justin B. surname: Searns fullname: Searns, Justin B. organization: Division of Pediatric Infectious Diseases, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA – sequence: 2 givenname: Christine C. surname: Robinson fullname: Robinson, Christine C. organization: Microbiology Department, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA – sequence: 3 givenname: Qi surname: Wei fullname: Wei, Qi organization: Microbiology Department, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA – sequence: 4 givenname: Ji surname: Yuan fullname: Yuan, Ji organization: Microbiology Department, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA – sequence: 5 givenname: Stacey surname: Hamilton fullname: Hamilton, Stacey organization: Microbiology Department, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA – sequence: 6 givenname: Kristin surname: Pretty fullname: Pretty, Kristin organization: Microbiology Department, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA – sequence: 7 givenname: Nathan surname: Donaldson fullname: Donaldson, Nathan organization: Department of Orthopedic Surgery, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA – sequence: 8 givenname: Sarah K. surname: Parker fullname: Parker, Sarah K. organization: Division of Pediatric Infectious Diseases, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA – sequence: 9 givenname: Samuel R. surname: Dominguez fullname: Dominguez, Samuel R. email: Samuel.dominguez@childrenscolorado.org organization: Division of Pediatric Infectious Diseases, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA
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Keywords	Osteomyelitis Infection Diagnostics NPV Septic arthritis PPV MDP qPCR Pediatric CHCO
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SubjectTerms	Anti-Bacterial Agents - therapeutic use antibiotic resistance antibiotic resistance genes Bacterial Proteins - genetics Child children clindamycin Clindamycin - therapeutic use Colorado Diagnostics Drug Resistance, Bacterial Female hospitals Humans Infection Kingella kingae Kingella kingae - genetics Kingella kingae - isolation & purification Male methicillin-resistant Staphylococcus aureus Methyltransferases - genetics microbial detection musculoskeletal system Neisseriaceae Infections - diagnosis Osteoarthritis - microbiology Osteomyelitis Osteomyelitis - microbiology patients Pediatric phenotype quantitative polymerase chain reaction Septic arthritis
Title	Validation of a novel molecular diagnostic panel for pediatric musculoskeletal infections: Integration of the Cepheid Xpert MRSA/SA SSTI and laboratory-developed real-time PCR assays for clindamycin resistance genes and Kingella kingae detection
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