Detection of hypervirulent Klebsiella pneumoniae (hvKp) strains directly from spiked blood cultures using a commercial Loop-Mediated isothermal amplification (LAMP) assay
Abstract Background Hypervirulent K. pneumoniae (hvKp) strains are characterized by their enhanced ability to evade immune responses and disseminate systemically. Rapid identification of hvKp strains is critical for guiding clinical management and implementing effective infection control measures. L...
Saved in:
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2025-08-01
|
| Series: | Annals of Clinical Microbiology and Antimicrobials |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12941-025-00817-4 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849226610454036480 |
|---|---|
| author | Domingo Fernández Vecilla Jorge Rodríguez Grande Nuria Fraile Valcárcel Zaira Moure García Sergio García Fernández María Siller Ruiz María Pía Roiz Mesones José Luis Díaz de Tuesta del Arco Mikel Joseba Urrutikoetxea Gutiérrez María Catalina Lomoro María Carmen Fariñas Alain Ocampo-Sosa |
| author_facet | Domingo Fernández Vecilla Jorge Rodríguez Grande Nuria Fraile Valcárcel Zaira Moure García Sergio García Fernández María Siller Ruiz María Pía Roiz Mesones José Luis Díaz de Tuesta del Arco Mikel Joseba Urrutikoetxea Gutiérrez María Catalina Lomoro María Carmen Fariñas Alain Ocampo-Sosa |
| author_sort | Domingo Fernández Vecilla |
| collection | DOAJ |
| description | Abstract Background Hypervirulent K. pneumoniae (hvKp) strains are characterized by their enhanced ability to evade immune responses and disseminate systemically. Rapid identification of hvKp strains is critical for guiding clinical management and implementing effective infection control measures. Loop-Mediated Isothermal Amplification (LAMP) assays provide a rapid and cost-effective method for detecting bacterial pathogens. This study evaluates the performance of the Eazyplex® hvKp assay for the direct detection of hvKp strains from spiked blood cultures. Methods We collected 20 K. pneumoniae (Kp) isolates between December 2021 and August 2024 from two hospitals in Northern Spain. Capsular serotyping and virulence gene detection were performed using PCR and whole-genome sequencing (WGS). The Eazyplex® hvKp LAMP assay was tested on spiked blood cultures inoculated with hvKp isolates. Virulence profiles were assessed using the Kleborate scoring system. Results Seventeen Kp isolates had a Kleborate score of ≥ 2, suggesting high virulence. The LAMP assay detected 87 out of 95 virulence targets, demonstrating an overall accuracy of 91.5%. Although eight target genes were not directly detected, fluorescence signals indicated amplification in all cases. The assay identified 16 strains with high virulence profiles (score ≥ 3), with 14 strains scoring 4 or 5. The LAMP-based test effectively detected hvKp directly from blood cultures, with time-to-results ranging from 6:43 to 17:11 min. Conclussion The EazyplexR hvKp LAMP assay is a rapid and effective method for identifying hvKp strains directly from blood cultures. This study supports its potential utility in clinical microbiology for early detection and epidemiological surveillance of hvKp infections. However, limitations in the Kleborate scoring system indicate that additional virulence biomarkers may be needed to improve the accuracy of hvKp classification. |
| format | Article |
| id | doaj-art-e192da7999aa4d13a35fffebae15e893 |
| institution | Kabale University |
| issn | 1476-0711 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Annals of Clinical Microbiology and Antimicrobials |
| spelling | doaj-art-e192da7999aa4d13a35fffebae15e8932025-08-24T11:12:41ZengBMCAnnals of Clinical Microbiology and Antimicrobials1476-07112025-08-012411710.1186/s12941-025-00817-4Detection of hypervirulent Klebsiella pneumoniae (hvKp) strains directly from spiked blood cultures using a commercial Loop-Mediated isothermal amplification (LAMP) assayDomingo Fernández Vecilla0Jorge Rodríguez Grande1Nuria Fraile Valcárcel2Zaira Moure García3Sergio García Fernández4María Siller Ruiz5María Pía Roiz Mesones6José Luis Díaz de Tuesta del Arco7Mikel Joseba Urrutikoetxea Gutiérrez8María Catalina Lomoro9María Carmen Fariñas10Alain Ocampo-Sosa11Microbiology Service, Valdecilla Research Institute (IDIVAL), University Hospital Marqués de ValdecillaMicrobiology Service, Valdecilla Research Institute (IDIVAL), University Hospital Marqués de ValdecillaMicrobiology Service, Valdecilla Research Institute (IDIVAL), University Hospital Marqués de ValdecillaMicrobiology Service, Valdecilla Research Institute (IDIVAL), University Hospital Marqués de ValdecillaMicrobiology Service, Valdecilla Research Institute (IDIVAL), University Hospital Marqués de ValdecillaMicrobiology Service, Valdecilla Research Institute (IDIVAL), University Hospital Marqués de ValdecillaMicrobiology Service, Valdecilla Research Institute (IDIVAL), University Hospital Marqués de ValdecillaMicrobiology service, Basurto University HosptitalMicrobiology service, Basurto University HosptitalMompía HospitalCIBERINFEC, Health Institute Carlos IIIMicrobiology Service, Valdecilla Research Institute (IDIVAL), University Hospital Marqués de ValdecillaAbstract Background Hypervirulent K. pneumoniae (hvKp) strains are characterized by their enhanced ability to evade immune responses and disseminate systemically. Rapid identification of hvKp strains is critical for guiding clinical management and implementing effective infection control measures. Loop-Mediated Isothermal Amplification (LAMP) assays provide a rapid and cost-effective method for detecting bacterial pathogens. This study evaluates the performance of the Eazyplex® hvKp assay for the direct detection of hvKp strains from spiked blood cultures. Methods We collected 20 K. pneumoniae (Kp) isolates between December 2021 and August 2024 from two hospitals in Northern Spain. Capsular serotyping and virulence gene detection were performed using PCR and whole-genome sequencing (WGS). The Eazyplex® hvKp LAMP assay was tested on spiked blood cultures inoculated with hvKp isolates. Virulence profiles were assessed using the Kleborate scoring system. Results Seventeen Kp isolates had a Kleborate score of ≥ 2, suggesting high virulence. The LAMP assay detected 87 out of 95 virulence targets, demonstrating an overall accuracy of 91.5%. Although eight target genes were not directly detected, fluorescence signals indicated amplification in all cases. The assay identified 16 strains with high virulence profiles (score ≥ 3), with 14 strains scoring 4 or 5. The LAMP-based test effectively detected hvKp directly from blood cultures, with time-to-results ranging from 6:43 to 17:11 min. Conclussion The EazyplexR hvKp LAMP assay is a rapid and effective method for identifying hvKp strains directly from blood cultures. This study supports its potential utility in clinical microbiology for early detection and epidemiological surveillance of hvKp infections. However, limitations in the Kleborate scoring system indicate that additional virulence biomarkers may be needed to improve the accuracy of hvKp classification.https://doi.org/10.1186/s12941-025-00817-4LAMPHypervirulentKlebsiella pneumoniaeDiagnosisSpiked blood cultures |
| spellingShingle | Domingo Fernández Vecilla Jorge Rodríguez Grande Nuria Fraile Valcárcel Zaira Moure García Sergio García Fernández María Siller Ruiz María Pía Roiz Mesones José Luis Díaz de Tuesta del Arco Mikel Joseba Urrutikoetxea Gutiérrez María Catalina Lomoro María Carmen Fariñas Alain Ocampo-Sosa Detection of hypervirulent Klebsiella pneumoniae (hvKp) strains directly from spiked blood cultures using a commercial Loop-Mediated isothermal amplification (LAMP) assay Annals of Clinical Microbiology and Antimicrobials LAMP Hypervirulent Klebsiella pneumoniae Diagnosis Spiked blood cultures |
| title | Detection of hypervirulent Klebsiella pneumoniae (hvKp) strains directly from spiked blood cultures using a commercial Loop-Mediated isothermal amplification (LAMP) assay |
| title_full | Detection of hypervirulent Klebsiella pneumoniae (hvKp) strains directly from spiked blood cultures using a commercial Loop-Mediated isothermal amplification (LAMP) assay |
| title_fullStr | Detection of hypervirulent Klebsiella pneumoniae (hvKp) strains directly from spiked blood cultures using a commercial Loop-Mediated isothermal amplification (LAMP) assay |
| title_full_unstemmed | Detection of hypervirulent Klebsiella pneumoniae (hvKp) strains directly from spiked blood cultures using a commercial Loop-Mediated isothermal amplification (LAMP) assay |
| title_short | Detection of hypervirulent Klebsiella pneumoniae (hvKp) strains directly from spiked blood cultures using a commercial Loop-Mediated isothermal amplification (LAMP) assay |
| title_sort | detection of hypervirulent klebsiella pneumoniae hvkp strains directly from spiked blood cultures using a commercial loop mediated isothermal amplification lamp assay |
| topic | LAMP Hypervirulent Klebsiella pneumoniae Diagnosis Spiked blood cultures |
| url | https://doi.org/10.1186/s12941-025-00817-4 |
| work_keys_str_mv | AT domingofernandezvecilla detectionofhypervirulentklebsiellapneumoniaehvkpstrainsdirectlyfromspikedbloodculturesusingacommercialloopmediatedisothermalamplificationlampassay AT jorgerodriguezgrande detectionofhypervirulentklebsiellapneumoniaehvkpstrainsdirectlyfromspikedbloodculturesusingacommercialloopmediatedisothermalamplificationlampassay AT nuriafrailevalcarcel detectionofhypervirulentklebsiellapneumoniaehvkpstrainsdirectlyfromspikedbloodculturesusingacommercialloopmediatedisothermalamplificationlampassay AT zairamouregarcia detectionofhypervirulentklebsiellapneumoniaehvkpstrainsdirectlyfromspikedbloodculturesusingacommercialloopmediatedisothermalamplificationlampassay AT sergiogarciafernandez detectionofhypervirulentklebsiellapneumoniaehvkpstrainsdirectlyfromspikedbloodculturesusingacommercialloopmediatedisothermalamplificationlampassay AT mariasillerruiz detectionofhypervirulentklebsiellapneumoniaehvkpstrainsdirectlyfromspikedbloodculturesusingacommercialloopmediatedisothermalamplificationlampassay AT mariapiaroizmesones detectionofhypervirulentklebsiellapneumoniaehvkpstrainsdirectlyfromspikedbloodculturesusingacommercialloopmediatedisothermalamplificationlampassay AT joseluisdiazdetuestadelarco detectionofhypervirulentklebsiellapneumoniaehvkpstrainsdirectlyfromspikedbloodculturesusingacommercialloopmediatedisothermalamplificationlampassay AT mikeljosebaurrutikoetxeagutierrez detectionofhypervirulentklebsiellapneumoniaehvkpstrainsdirectlyfromspikedbloodculturesusingacommercialloopmediatedisothermalamplificationlampassay AT mariacatalinalomoro detectionofhypervirulentklebsiellapneumoniaehvkpstrainsdirectlyfromspikedbloodculturesusingacommercialloopmediatedisothermalamplificationlampassay AT mariacarmenfarinas detectionofhypervirulentklebsiellapneumoniaehvkpstrainsdirectlyfromspikedbloodculturesusingacommercialloopmediatedisothermalamplificationlampassay AT alainocampososa detectionofhypervirulentklebsiellapneumoniaehvkpstrainsdirectlyfromspikedbloodculturesusingacommercialloopmediatedisothermalamplificationlampassay |