Simple and specific detection of <i>Bordetella holmesii</i> by using a loop‐mediated isothermal amplification assay

Otsuka, Nao; Yoshino, Shuji; Kawano, Keiichi; Toyoizumi-Ajisaka, Hiromi; Shibayama, Keigo; Kamachi, Kazunari

doi:10.1111/j.1348-0421.2012.00465.x

Cited by 8 publications

(4 citation statements)

References 19 publications

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“…There are reports proving that PRN (−) strains do not cause stronger clinical symptoms in infants less than 6 months old, from whom they were isolated [ 17 , 19 ]. However, there are also reports demonstrating that PRN (−) isolates show greater virulence in populations immunised with aP vaccines [ 65 ]. This observation has also been suggested recently by US researchers [ 40 ].…”

Section: Discussionmentioning

confidence: 99%

Effectiveness of experimental and commercial pertussis vaccines in the elimination of Bordetella pertussis isolates with different genetic profiles in murine model

Prygiel

Mosiej

Wdowiak

et al. 2021

Med Microbiol Immunol

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The aim of this study was to compare the elimination of Bordetella pertussis clinical isolates, representing different genotypes in relation to alleles encoding virulence factors (MLST—multi-locus antigen sequence typing), MLVA type (multi-locus variable-number tandem repeat analysis) and PFGE group (pulsed-field gel electrophoresis) from the lungs of naive mice or mice were immunised with the commercial whole-cell pertussis vaccine, the acellular pertussis vaccine and the experimental whole-cell pertussis vaccine. Molecular data indicate that the resurgence of pertussis in populations with high vaccine coverage is associated with genomic adaptation of B. pertussis, to vaccine selection pressure. Pertactin-negative B. pertussis isolates were suspected to contribute to the reduced vaccine effectiveness. It was shown that one of the isolates used is PRN deficient. The mice were intranasally challenged with bacterial suspension containing approximately 5 × 10 7 CFU/ml B. pertussis. The immunogenicity of the tested vaccines against PT (pertussis toxin), PRN (pertactin), FHA (filamentous haemagglutinin) and FIM (fimbriae types 2 and 3) was examined. The commercial whole-cell and acellular pertussis vaccines induced an immunity effective at eliminating the genetically different B. pertussis isolates from the lungs. However, the elimination of the PRN-deficient isolate from the lungs of mice vaccinated with commercial vaccines was delayed as compared to the PRN ( +) isolate, suggesting phenotypic differences with the circulating isolates and vaccine strains. The most effective vaccine was the experimental vaccine with the composition identical to that of the strains used for infection.

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Section: Discussionmentioning

confidence: 99%

Effectiveness of experimental and commercial pertussis vaccines in the elimination of Bordetella pertussis isolates with different genetic profiles in murine model

Prygiel

Mosiej

Wdowiak

et al. 2021

Med Microbiol Immunol

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“…Other methods that can be used to avoid misidentification is the recA gene amplification using a real-time polymerase chain reaction (PCR) or a loop-mediated isothermal amplification (LAMP). 9 At the moment, the European Committee on Antimicrobial Susceptibility Testing (EUCAST) do not have standardized breakpoints for B. holmesii. The AST analysis was performed by the disk-diffusion method using the interpretation values of the inhibition halo for Enterobacteriaceae and MIC values were calculated using E-test.…”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Prosthetic joint infection by Bordetella holmesii: Case report and a review of the literature

et al. 2020

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Introduction: Bordetella holmesii is a Gram-negative coccobacillus involved in different infections mostly described in case reports. Prosthetic joint infections in relation to this pathogen are rare. Here, we present the third case of B. holmesii in a patient without anatomical or functional spleen dysfunction. Case report: The patient was a 62-year-old female with a total knee prosthesis implanted in 1997 that required multiple replacements of the femoral component due to aseptic loosening in the past years. The patient was admitted to our hospital for an elective replacement surgery due to new radiological signs of loosening. B. holmesii was isolated from synovial fluid obtained during surgery. The identification was performed by matrix-assisted laser desorption ionization–time of flight mass spectrometry and confirmed by 16S rRNA gene amplification and sequencing. Antibiotic treatment was started but 14 days after surgery the patient presented pain and joint effusion. An arthrocentesis was performed and synovial fluid culture was positive again for B. holmesii. Surgical debridement including polyethylene replacement was performed and antibiotic treatment was continued for 3 months. After a 2-year follow-up period, the patient remained asymptomatic and physical examination showed normal function of the prosthesis. Conclusion: B. holmesii is an uncommon cause of bone and joint infections. This case indicates that this microorganism is a potential pathogen of prosthetic or native arthritis, and it should be considered when cultures are negative and in cases presenting torpid evolution.

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“…In the LAMP assays for detecting bacteria responsible for endocarditis, bacteria from the samples had to be previously cultured and DNA had to be isolated from a pure culture [49, 50]. In other studies, bacteria (including the causative agents of endocarditis, but not E. hirae ) were used directly in the LAMP test without prior culturing [48, 51, 52]. In this study, E. hirae were detected in clinical isolates and heart tissue samples for the first time with the use of the LAMP assay.…”

Section: Discussionmentioning

confidence: 99%

The application of the loop-mediated isothermal amplification (LAMP) method for diagnosing Enterococcus hirae-associated endocarditis outbreaks in chickens

2019

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Background Enterococcus hirae is considered a part of the normal intestinal biota of several domestic animals, including poultry. However, this species is also associated with infective endocarditis in chickens, a disease that leads to unexpected deaths and serious economical losses. Enterococcus hirae is identified predominantly with the use of conventional bacteriological methods, biochemical tests and PCR. Rapid, sensitive and specific methods for detecting E. hirae in clinical samples are required in poultry production. The aim of this study was to use the Loop-Mediated Isothermal Amplification (LAMP) for the identification and quantification of E. hirae in heart samples from broiler chickens. Results The specificity of the LAMP method was confirmed for 7 enterococcal strains and 3 non-enterococcal strains. E. hirae was detected in all of the 22 analyzed clinical bacterial isolates and in all of the 9 heart samples. Three sets of primers supported the detection of E. hirae with high sensitivity and specificity within one hour. The highest detection rate of a LAMP product was approximately 7 min for an E. hirae strain and 12 min for a positive heart sample. The detection limit for the E. hirae ATCC 10541 standard was 1.3 × 10 2 CFU (43.4 fg) or 13.8 copies of the E. hirae genome equivalent per reaction. The reaction was 10-fold more sensitive than conventional species-specific PCR. The LAMP assay supported the determination of the E. hirae load in chicken hearts with endocarditis in field cases. The average number of E. hirae cells in hearts was 5.19 × 10 7 CFU/g of tissue, and the average number of E. hirae genome equivalents in hearts was 5.51× 10 6 copies/g of tissue. Bacterial counts were significantly higher in the LAMP assay than in the standard plate count. Conclusions The LAMP assay is a useful diagnostic tool and an effective alternative to conventional methods for the detection of this enterococcal species. The sod A-based LAMP assay supported direct identification of E. hirae from pure cultures and heart samples without previous bacterial cultivation. This is the first study to apply the LAMP method for the purpose of diagnosing E. hirae- associated endocarditis in poultry.

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Simple and specific detection of Bordetella holmesii by using a loop‐mediated isothermal amplification assay

Cited by 8 publications

References 19 publications

Effectiveness of experimental and commercial pertussis vaccines in the elimination of Bordetella pertussis isolates with different genetic profiles in murine model

Effectiveness of experimental and commercial pertussis vaccines in the elimination of Bordetella pertussis isolates with different genetic profiles in murine model

Prosthetic joint infection by Bordetella holmesii: Case report and a review of the literature

The application of the loop-mediated isothermal amplification (LAMP) method for diagnosing Enterococcus hirae-associated endocarditis outbreaks in chickens

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