Clinical validation of a multiplex droplet digital PCR for diagnosing suspected bloodstream infections in ICU practice: a promising diagnostic tool
Abstract Background Droplet digital PCR (ddPCR) has emerged as a promising tool of pathogen detection in bloodstream infections (BSIs) in critical care medicine. However, different ddPCR platforms have variable sensitivity and specificity for diverse microorganisms at various infection sites. There...
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| Format: | Article |
| Language: | English |
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BMC
2022-08-01
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| Series: | Critical Care |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13054-022-04116-8 |
| _version_ | 1828183311195832320 |
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| author | Jing Wu Bin Tang Yuzhen Qiu Ruoming Tan Jialin Liu Jiang Xia Jing Zhang Jingjing Huang Jieming Qu Jingyong Sun Xiaoli Wang Hongping Qu |
| author_facet | Jing Wu Bin Tang Yuzhen Qiu Ruoming Tan Jialin Liu Jiang Xia Jing Zhang Jingjing Huang Jieming Qu Jingyong Sun Xiaoli Wang Hongping Qu |
| author_sort | Jing Wu |
| collection | DOAJ |
| description | Abstract Background Droplet digital PCR (ddPCR) has emerged as a promising tool of pathogen detection in bloodstream infections (BSIs) in critical care medicine. However, different ddPCR platforms have variable sensitivity and specificity for diverse microorganisms at various infection sites. There is still a lack of prospective clinical studies aimed at validating and interpreting the discrepant ddPCR results for diagnosing BSI in intensive care unit (ICU) practice. Methods A prospective diagnostic study of multiplex ddPCR panels was conducted in a general ICU from May 21, 2021, to December 22, 2021. Paired blood cultures (BCs) and ddPCRs (2.5 h) were obtained synchronously to detect the 12 most common BSI pathogens and three antimicrobial resistance (AMR) genes. Firstly, ddPCR performance was compared to definite BSI. Secondly, clinical validation of ddPCR was compared to composite clinical diagnosis. Sensitivity, specificity, and positive and negative predictive values were calculated. Thirdly, the positive rate of AMR genes and related analysis was presented. Results A total of 438 episodes of suspected BSIs occurring in 150 critical patients were enrolled. BC and ddPCR were positive for targeted bacteria in 40 (9.1%) and 180 (41.1%) cases, respectively. There were 280 concordant and 158 discordant. In comparison with BCs, the sensitivity of ddPCR ranged from 58.8 to 86.7% with an aggregate of 72.5% in different species, with corresponding specificity ranging from 73.5 to 92.2% with an aggregate of 63.1%. Furthermore, the rate of ddPCR+/BC− results was 33.6% (147/438) with 87.1% (128 of 147) cases was associated with probable (n = 108) or possible (n = 20) BSIs. When clinically diagnosed BSI was used as true positive, the final sensitivity and specificity of ddPCR increased to 84.9% and 92.5%, respectively. In addition, 40 bla KPC, 3bla NDM, and 38 mecA genes were detected, among which 90.5% were definitely positive for bla KPC. Further, 65.8% specimens were predicted to be mecA-positive in Staphylococcus sp. according to all microbiological analysis. Conclusions The multiplexed ddPCR is a flexible and universal platform, which can be used as an add-on complementary to conventional BC. When combined with clinical infection evidence, ddPCR shows potential advantages for rapidly diagnosing suspected BSIs and AMR genes in ICU practice. |
| first_indexed | 2024-04-12T06:30:29Z |
| format | Article |
| id | doaj.art-ae9bafa603674d7f8cc3be5f2d376d77 |
| institution | Directory Open Access Journal |
| issn | 1364-8535 |
| language | English |
| last_indexed | 2024-04-12T06:30:29Z |
| publishDate | 2022-08-01 |
| publisher | BMC |
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| series | Critical Care |
| spelling | doaj.art-ae9bafa603674d7f8cc3be5f2d376d772022-12-22T03:44:01ZengBMCCritical Care1364-85352022-08-0126111110.1186/s13054-022-04116-8Clinical validation of a multiplex droplet digital PCR for diagnosing suspected bloodstream infections in ICU practice: a promising diagnostic toolJing Wu0Bin Tang1Yuzhen Qiu2Ruoming Tan3Jialin Liu4Jiang Xia5Jing Zhang6Jingjing Huang7Jieming Qu8Jingyong Sun9Xiaoli Wang10Hongping Qu11Department of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of MedicineDepartment of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of MedicineDepartment of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of MedicineDepartment of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of MedicineDepartment of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of MedicinePilot Gene Technologies (Hangzhou) Co., LtdPilot Gene Technologies (Hangzhou) Co., LtdDepartment of Pharmacy, Ruijin Hospital, Shanghai Jiao Tong University School of MedicineDepartment of Pulmonary and Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of MedicineDepartment of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of MedicineDepartment of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of MedicineDepartment of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of MedicineAbstract Background Droplet digital PCR (ddPCR) has emerged as a promising tool of pathogen detection in bloodstream infections (BSIs) in critical care medicine. However, different ddPCR platforms have variable sensitivity and specificity for diverse microorganisms at various infection sites. There is still a lack of prospective clinical studies aimed at validating and interpreting the discrepant ddPCR results for diagnosing BSI in intensive care unit (ICU) practice. Methods A prospective diagnostic study of multiplex ddPCR panels was conducted in a general ICU from May 21, 2021, to December 22, 2021. Paired blood cultures (BCs) and ddPCRs (2.5 h) were obtained synchronously to detect the 12 most common BSI pathogens and three antimicrobial resistance (AMR) genes. Firstly, ddPCR performance was compared to definite BSI. Secondly, clinical validation of ddPCR was compared to composite clinical diagnosis. Sensitivity, specificity, and positive and negative predictive values were calculated. Thirdly, the positive rate of AMR genes and related analysis was presented. Results A total of 438 episodes of suspected BSIs occurring in 150 critical patients were enrolled. BC and ddPCR were positive for targeted bacteria in 40 (9.1%) and 180 (41.1%) cases, respectively. There were 280 concordant and 158 discordant. In comparison with BCs, the sensitivity of ddPCR ranged from 58.8 to 86.7% with an aggregate of 72.5% in different species, with corresponding specificity ranging from 73.5 to 92.2% with an aggregate of 63.1%. Furthermore, the rate of ddPCR+/BC− results was 33.6% (147/438) with 87.1% (128 of 147) cases was associated with probable (n = 108) or possible (n = 20) BSIs. When clinically diagnosed BSI was used as true positive, the final sensitivity and specificity of ddPCR increased to 84.9% and 92.5%, respectively. In addition, 40 bla KPC, 3bla NDM, and 38 mecA genes were detected, among which 90.5% were definitely positive for bla KPC. Further, 65.8% specimens were predicted to be mecA-positive in Staphylococcus sp. according to all microbiological analysis. Conclusions The multiplexed ddPCR is a flexible and universal platform, which can be used as an add-on complementary to conventional BC. When combined with clinical infection evidence, ddPCR shows potential advantages for rapidly diagnosing suspected BSIs and AMR genes in ICU practice.https://doi.org/10.1186/s13054-022-04116-8Clinical validationDroplet digital PCRBloodstream infections (BSIs)Critically ill patientsDiscordant results |
| spellingShingle | Jing Wu Bin Tang Yuzhen Qiu Ruoming Tan Jialin Liu Jiang Xia Jing Zhang Jingjing Huang Jieming Qu Jingyong Sun Xiaoli Wang Hongping Qu Clinical validation of a multiplex droplet digital PCR for diagnosing suspected bloodstream infections in ICU practice: a promising diagnostic tool Critical Care Clinical validation Droplet digital PCR Bloodstream infections (BSIs) Critically ill patients Discordant results |
| title | Clinical validation of a multiplex droplet digital PCR for diagnosing suspected bloodstream infections in ICU practice: a promising diagnostic tool |
| title_full | Clinical validation of a multiplex droplet digital PCR for diagnosing suspected bloodstream infections in ICU practice: a promising diagnostic tool |
| title_fullStr | Clinical validation of a multiplex droplet digital PCR for diagnosing suspected bloodstream infections in ICU practice: a promising diagnostic tool |
| title_full_unstemmed | Clinical validation of a multiplex droplet digital PCR for diagnosing suspected bloodstream infections in ICU practice: a promising diagnostic tool |
| title_short | Clinical validation of a multiplex droplet digital PCR for diagnosing suspected bloodstream infections in ICU practice: a promising diagnostic tool |
| title_sort | clinical validation of a multiplex droplet digital pcr for diagnosing suspected bloodstream infections in icu practice a promising diagnostic tool |
| topic | Clinical validation Droplet digital PCR Bloodstream infections (BSIs) Critically ill patients Discordant results |
| url | https://doi.org/10.1186/s13054-022-04116-8 |
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