Detecting the Presence of Bacterial DNA and RNA by Polymerase Chain Reaction to Diagnose Suspected Periprosthetic Joint Infection after Antibiotic Therapy

Objective To explore the diagnostic efficiency of DNA‐based and RNA‐based quantitative polymerase chain reaction (qPCR) analyses for periprosthetic joint infection (PJI). Methods To determine the detection limit of DNA‐based and RNA‐based qPCR in vitro, Staphylococcus aureus and Escherichia coli str...

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Veröffentlicht in:Orthopaedic surgery 2018-02, Vol.10 (1), p.40-46
Hauptverfasser: Fang, Xin‐yu, Li, Wen‐bo, Zhang, Chao‐fan, Huang, Zi‐da, Zeng, Hui‐yi, Dong, Zheng, Zhang, Wen‐ming
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container_end_page 46
container_issue 1
container_start_page 40
container_title Orthopaedic surgery
container_volume 10
creator Fang, Xin‐yu
Li, Wen‐bo
Zhang, Chao‐fan
Huang, Zi‐da
Zeng, Hui‐yi
Dong, Zheng
Zhang, Wen‐ming
description Objective To explore the diagnostic efficiency of DNA‐based and RNA‐based quantitative polymerase chain reaction (qPCR) analyses for periprosthetic joint infection (PJI). Methods To determine the detection limit of DNA‐based and RNA‐based qPCR in vitro, Staphylococcus aureus and Escherichia coli strains were added to sterile synovial fluid obtained from a patient with knee osteoarthritis. Serial dilutions of samples were analyzed by DNA‐based and RNA‐based qPCR. Clinically, patients who were suspected of having PJI and eventually underwent revision arthroplasty in our hospital from July 2014 to December 2016 were screened. Preoperative puncture or intraoperative collection was performed on patients who met the inclusion and exclusion criteria to obtain synovial fluid. DNA‐based and RNA‐based PCR analyses and culture were performed on each synovial fluid sample. The patients’ demographic characteristics, medical history, and laboratory test results were recorded. The diagnostic efficiency of both PCR assays was compared with culture methods. Results The in vitro analysis demonstrated that DNA‐based qPCR assay was highly sensitive, with the detection limit being 1200 colony forming units (CFU)/mL of S. aureus and 3200 CFU/mL of E. coli. Meanwhile, The RNA‐based qPCR assay could detect 2300 CFU/mL of S. aureus and 11 000 CFU/mL of E. coli. Clinically, the sensitivity, specificity, and accuracy were 65.7%, 100%, and 81.6%, respectively, for the culture method; 81.5%, 84.8%, and 83.1%, respectively, for DNA‐based qPCR; and 73.6%, 100%, and 85.9%, respectively, for RNA‐based qPCR. Conclusions DNA‐based qPCR could detect suspected PJI with high sensitivity after antibiotic therapy. RNA‐based qPCR could reduce the false positive rates of DNA‐based assays. qPCR‐based methods could improve the efficiency of PJI diagnosis.
doi_str_mv 10.1111/os.12359
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Methods To determine the detection limit of DNA‐based and RNA‐based qPCR in vitro, Staphylococcus aureus and Escherichia coli strains were added to sterile synovial fluid obtained from a patient with knee osteoarthritis. Serial dilutions of samples were analyzed by DNA‐based and RNA‐based qPCR. Clinically, patients who were suspected of having PJI and eventually underwent revision arthroplasty in our hospital from July 2014 to December 2016 were screened. Preoperative puncture or intraoperative collection was performed on patients who met the inclusion and exclusion criteria to obtain synovial fluid. DNA‐based and RNA‐based PCR analyses and culture were performed on each synovial fluid sample. The patients’ demographic characteristics, medical history, and laboratory test results were recorded. The diagnostic efficiency of both PCR assays was compared with culture methods. Results The in vitro analysis demonstrated that DNA‐based qPCR assay was highly sensitive, with the detection limit being 1200 colony forming units (CFU)/mL of S. aureus and 3200 CFU/mL of E. coli. Meanwhile, The RNA‐based qPCR assay could detect 2300 CFU/mL of S. aureus and 11 000 CFU/mL of E. coli. Clinically, the sensitivity, specificity, and accuracy were 65.7%, 100%, and 81.6%, respectively, for the culture method; 81.5%, 84.8%, and 83.1%, respectively, for DNA‐based qPCR; and 73.6%, 100%, and 85.9%, respectively, for RNA‐based qPCR. Conclusions DNA‐based qPCR could detect suspected PJI with high sensitivity after antibiotic therapy. RNA‐based qPCR could reduce the false positive rates of DNA‐based assays. qPCR‐based methods could improve the efficiency of PJI diagnosis.</description><identifier>ISSN: 1757-7853</identifier><identifier>EISSN: 1757-7861</identifier><identifier>DOI: 10.1111/os.12359</identifier><identifier>PMID: 29383856</identifier><language>eng</language><publisher>Melbourne: John Wiley &amp; Sons Australia, Ltd</publisher><subject>Aged ; Anti-Bacterial Agents - therapeutic use ; Antibiotic therapy ; Antibiotics ; Arthroplasty, Replacement, Knee ; Bacterial DNA ; Bacterial RNA ; Basic Research ; Deoxyribonucleic acid ; DNA ; DNA, Bacterial - analysis ; E coli ; Efficiency ; Escherichia coli - isolation &amp; purification ; Escherichia coli Infections - diagnosis ; Female ; Humans ; Joint surgery ; Knee Prosthesis - adverse effects ; Male ; Middle Aged ; Osteoarthritis, Knee - surgery ; Periprosthetic joint infection ; Polymerase chain reaction ; Polymerase Chain Reaction - methods ; Postoperative Care - methods ; Prosthesis-Related Infections - diagnosis ; Prosthesis-Related Infections - microbiology ; Real‐time PCR ; RNA, Bacterial - analysis ; Sensitivity and Specificity ; Staphylococcal Infections - diagnosis ; Staphylococcus aureus - isolation &amp; purification ; Synovial Fluid - microbiology</subject><ispartof>Orthopaedic surgery, 2018-02, Vol.10 (1), p.40-46</ispartof><rights>2018 Chinese Orthopaedic Association and John Wiley &amp; Sons Australia, Ltd</rights><rights>2018 Chinese Orthopaedic Association and John Wiley &amp; Sons Australia, Ltd.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4059-5a6b633f9c1692f848a37f2ad35edcfe8e884bdd9e0bebfd2f5769a07323bbaf3</citedby><cites>FETCH-LOGICAL-c4059-5a6b633f9c1692f848a37f2ad35edcfe8e884bdd9e0bebfd2f5769a07323bbaf3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6594519/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6594519/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,724,777,781,882,1412,11543,27905,27906,45555,45556,46033,46457,53772,53774</link.rule.ids><linktorsrc>$$Uhttps://onlinelibrary.wiley.com/doi/abs/10.1111%2Fos.12359$$EView_record_in_Wiley-Blackwell$$FView_record_in_$$GWiley-Blackwell</linktorsrc><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29383856$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Fang, Xin‐yu</creatorcontrib><creatorcontrib>Li, Wen‐bo</creatorcontrib><creatorcontrib>Zhang, Chao‐fan</creatorcontrib><creatorcontrib>Huang, Zi‐da</creatorcontrib><creatorcontrib>Zeng, Hui‐yi</creatorcontrib><creatorcontrib>Dong, Zheng</creatorcontrib><creatorcontrib>Zhang, Wen‐ming</creatorcontrib><title>Detecting the Presence of Bacterial DNA and RNA by Polymerase Chain Reaction to Diagnose Suspected Periprosthetic Joint Infection after Antibiotic Therapy</title><title>Orthopaedic surgery</title><addtitle>Orthop Surg</addtitle><description>Objective To explore the diagnostic efficiency of DNA‐based and RNA‐based quantitative polymerase chain reaction (qPCR) analyses for periprosthetic joint infection (PJI). Methods To determine the detection limit of DNA‐based and RNA‐based qPCR in vitro, Staphylococcus aureus and Escherichia coli strains were added to sterile synovial fluid obtained from a patient with knee osteoarthritis. Serial dilutions of samples were analyzed by DNA‐based and RNA‐based qPCR. Clinically, patients who were suspected of having PJI and eventually underwent revision arthroplasty in our hospital from July 2014 to December 2016 were screened. Preoperative puncture or intraoperative collection was performed on patients who met the inclusion and exclusion criteria to obtain synovial fluid. DNA‐based and RNA‐based PCR analyses and culture were performed on each synovial fluid sample. The patients’ demographic characteristics, medical history, and laboratory test results were recorded. The diagnostic efficiency of both PCR assays was compared with culture methods. Results The in vitro analysis demonstrated that DNA‐based qPCR assay was highly sensitive, with the detection limit being 1200 colony forming units (CFU)/mL of S. aureus and 3200 CFU/mL of E. coli. Meanwhile, The RNA‐based qPCR assay could detect 2300 CFU/mL of S. aureus and 11 000 CFU/mL of E. coli. Clinically, the sensitivity, specificity, and accuracy were 65.7%, 100%, and 81.6%, respectively, for the culture method; 81.5%, 84.8%, and 83.1%, respectively, for DNA‐based qPCR; and 73.6%, 100%, and 85.9%, respectively, for RNA‐based qPCR. Conclusions DNA‐based qPCR could detect suspected PJI with high sensitivity after antibiotic therapy. RNA‐based qPCR could reduce the false positive rates of DNA‐based assays. qPCR‐based methods could improve the efficiency of PJI diagnosis.</description><subject>Aged</subject><subject>Anti-Bacterial Agents - therapeutic use</subject><subject>Antibiotic therapy</subject><subject>Antibiotics</subject><subject>Arthroplasty, Replacement, Knee</subject><subject>Bacterial DNA</subject><subject>Bacterial RNA</subject><subject>Basic Research</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA, Bacterial - analysis</subject><subject>E coli</subject><subject>Efficiency</subject><subject>Escherichia coli - isolation &amp; purification</subject><subject>Escherichia coli Infections - diagnosis</subject><subject>Female</subject><subject>Humans</subject><subject>Joint surgery</subject><subject>Knee Prosthesis - adverse effects</subject><subject>Male</subject><subject>Middle Aged</subject><subject>Osteoarthritis, Knee - surgery</subject><subject>Periprosthetic joint infection</subject><subject>Polymerase chain reaction</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Postoperative Care - methods</subject><subject>Prosthesis-Related Infections - diagnosis</subject><subject>Prosthesis-Related Infections - microbiology</subject><subject>Real‐time PCR</subject><subject>RNA, Bacterial - analysis</subject><subject>Sensitivity and Specificity</subject><subject>Staphylococcal Infections - diagnosis</subject><subject>Staphylococcus aureus - isolation &amp; purification</subject><subject>Synovial Fluid - microbiology</subject><issn>1757-7853</issn><issn>1757-7861</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kc1u1DAQgC0Eoj8g8QTIEhcuKXEcJ_YFadktUFTRVVvOlp2Md11l7dT2gvIqPC0uKSs44MtYmm--Gc0g9IqUZyS_dz6ekYoy8QQdk5a1Rcsb8vTwZ_QIncR4V5aNoG37HB1VgnLKWXOMfq4gQZes2-C0BbwOEMF1gL3BH1SXIFg14NXXBVaux9c56gmv_TDtIKgIeLlV1uFryKj1DiePV1ZtnM-pm30csxl6vM6WMfiYGyTb4S_euoQvnIG5SJncBi9cstr6B-B2m-Xj9AI9M2qI8PIxnqJvH89vl5-Ly6tPF8vFZdHVJRMFU41uKDWiI42oDK-5oq2pVE8Z9J0BDpzXuu8FlBq06SvD2kaosqUV1VoZeorez95xr3e5BFwKapBjsDsVJumVlf9mnN3Kjf8uGyZqRkQWvHkUBH-_h5jknd8Hl2eWVVly0lBek0y9nakuryIGMIcOpJQPV5Q-yt9XzOjrvyc6gH_OloFiBn7YAab_iuTVzSz8BVHdqZk</recordid><startdate>201802</startdate><enddate>201802</enddate><creator>Fang, Xin‐yu</creator><creator>Li, Wen‐bo</creator><creator>Zhang, Chao‐fan</creator><creator>Huang, Zi‐da</creator><creator>Zeng, Hui‐yi</creator><creator>Dong, Zheng</creator><creator>Zhang, Wen‐ming</creator><general>John Wiley &amp; Sons Australia, Ltd</general><general>John Wiley &amp; Sons, Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>K9.</scope><scope>5PM</scope></search><sort><creationdate>201802</creationdate><title>Detecting the Presence of Bacterial DNA and RNA by Polymerase Chain Reaction to Diagnose Suspected Periprosthetic Joint Infection after Antibiotic Therapy</title><author>Fang, Xin‐yu ; Li, Wen‐bo ; Zhang, Chao‐fan ; Huang, Zi‐da ; Zeng, Hui‐yi ; Dong, Zheng ; Zhang, Wen‐ming</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4059-5a6b633f9c1692f848a37f2ad35edcfe8e884bdd9e0bebfd2f5769a07323bbaf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Aged</topic><topic>Anti-Bacterial Agents - therapeutic use</topic><topic>Antibiotic therapy</topic><topic>Antibiotics</topic><topic>Arthroplasty, Replacement, Knee</topic><topic>Bacterial DNA</topic><topic>Bacterial RNA</topic><topic>Basic Research</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA, Bacterial - analysis</topic><topic>E coli</topic><topic>Efficiency</topic><topic>Escherichia coli - isolation &amp; purification</topic><topic>Escherichia coli Infections - diagnosis</topic><topic>Female</topic><topic>Humans</topic><topic>Joint surgery</topic><topic>Knee Prosthesis - adverse effects</topic><topic>Male</topic><topic>Middle Aged</topic><topic>Osteoarthritis, Knee - surgery</topic><topic>Periprosthetic joint infection</topic><topic>Polymerase chain reaction</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Postoperative Care - methods</topic><topic>Prosthesis-Related Infections - diagnosis</topic><topic>Prosthesis-Related Infections - microbiology</topic><topic>Real‐time PCR</topic><topic>RNA, Bacterial - analysis</topic><topic>Sensitivity and Specificity</topic><topic>Staphylococcal Infections - diagnosis</topic><topic>Staphylococcus aureus - isolation &amp; purification</topic><topic>Synovial Fluid - microbiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Fang, Xin‐yu</creatorcontrib><creatorcontrib>Li, Wen‐bo</creatorcontrib><creatorcontrib>Zhang, Chao‐fan</creatorcontrib><creatorcontrib>Huang, Zi‐da</creatorcontrib><creatorcontrib>Zeng, Hui‐yi</creatorcontrib><creatorcontrib>Dong, Zheng</creatorcontrib><creatorcontrib>Zhang, Wen‐ming</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Health &amp; Medical Complete (Alumni)</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Orthopaedic surgery</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext_linktorsrc</fulltext></delivery><addata><au>Fang, Xin‐yu</au><au>Li, Wen‐bo</au><au>Zhang, Chao‐fan</au><au>Huang, Zi‐da</au><au>Zeng, Hui‐yi</au><au>Dong, Zheng</au><au>Zhang, Wen‐ming</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detecting the Presence of Bacterial DNA and RNA by Polymerase Chain Reaction to Diagnose Suspected Periprosthetic Joint Infection after Antibiotic Therapy</atitle><jtitle>Orthopaedic surgery</jtitle><addtitle>Orthop Surg</addtitle><date>2018-02</date><risdate>2018</risdate><volume>10</volume><issue>1</issue><spage>40</spage><epage>46</epage><pages>40-46</pages><issn>1757-7853</issn><eissn>1757-7861</eissn><abstract>Objective To explore the diagnostic efficiency of DNA‐based and RNA‐based quantitative polymerase chain reaction (qPCR) analyses for periprosthetic joint infection (PJI). Methods To determine the detection limit of DNA‐based and RNA‐based qPCR in vitro, Staphylococcus aureus and Escherichia coli strains were added to sterile synovial fluid obtained from a patient with knee osteoarthritis. Serial dilutions of samples were analyzed by DNA‐based and RNA‐based qPCR. Clinically, patients who were suspected of having PJI and eventually underwent revision arthroplasty in our hospital from July 2014 to December 2016 were screened. Preoperative puncture or intraoperative collection was performed on patients who met the inclusion and exclusion criteria to obtain synovial fluid. DNA‐based and RNA‐based PCR analyses and culture were performed on each synovial fluid sample. The patients’ demographic characteristics, medical history, and laboratory test results were recorded. The diagnostic efficiency of both PCR assays was compared with culture methods. Results The in vitro analysis demonstrated that DNA‐based qPCR assay was highly sensitive, with the detection limit being 1200 colony forming units (CFU)/mL of S. aureus and 3200 CFU/mL of E. coli. Meanwhile, The RNA‐based qPCR assay could detect 2300 CFU/mL of S. aureus and 11 000 CFU/mL of E. coli. Clinically, the sensitivity, specificity, and accuracy were 65.7%, 100%, and 81.6%, respectively, for the culture method; 81.5%, 84.8%, and 83.1%, respectively, for DNA‐based qPCR; and 73.6%, 100%, and 85.9%, respectively, for RNA‐based qPCR. Conclusions DNA‐based qPCR could detect suspected PJI with high sensitivity after antibiotic therapy. RNA‐based qPCR could reduce the false positive rates of DNA‐based assays. qPCR‐based methods could improve the efficiency of PJI diagnosis.</abstract><cop>Melbourne</cop><pub>John Wiley &amp; Sons Australia, Ltd</pub><pmid>29383856</pmid><doi>10.1111/os.12359</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record>
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subjects Aged
Anti-Bacterial Agents - therapeutic use
Antibiotic therapy
Antibiotics
Arthroplasty, Replacement, Knee
Bacterial DNA
Bacterial RNA
Basic Research
Deoxyribonucleic acid
DNA
DNA, Bacterial - analysis
E coli
Efficiency
Escherichia coli - isolation & purification
Escherichia coli Infections - diagnosis
Female
Humans
Joint surgery
Knee Prosthesis - adverse effects
Male
Middle Aged
Osteoarthritis, Knee - surgery
Periprosthetic joint infection
Polymerase chain reaction
Polymerase Chain Reaction - methods
Postoperative Care - methods
Prosthesis-Related Infections - diagnosis
Prosthesis-Related Infections - microbiology
Real‐time PCR
RNA, Bacterial - analysis
Sensitivity and Specificity
Staphylococcal Infections - diagnosis
Staphylococcus aureus - isolation & purification
Synovial Fluid - microbiology
title Detecting the Presence of Bacterial DNA and RNA by Polymerase Chain Reaction to Diagnose Suspected Periprosthetic Joint Infection after Antibiotic Therapy
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