Eubacterial approach to the diagnosis of bacterial infection

The aim was to evaluate "16S rDNA PCR and sequencing" (PCR) for identification of bacterial DNA in heart valves in routine diagnosis of infective endocarditis (IE). Heart valves from 74 patients with suspected infective endocarditis, and 16 controls were analysed by histology, culture and PCR. Results from blood culture served as the gold standard. Patients were classified according to the Duke criteria. The final classification resulted in 57 definitive cases of IE, 7 possible, and 10 cases without IE. Sensitivity of valve culture was 26% and specificity 62%. Sensitivity of PCR was 72% and specificity 100%. In patients who had received antibiotic treatment for less than 5 days before surgery, sensitivity of culture and PCR were comparable. In patients who had received antibiotic treatment for more than 5 days, sensitivity of valve culture was markedly reduced compared to sensitivity of PCR. In three of seven blood-culture-negative cases PCR was positive, including two cases with non-cultivable bacteria. No PCR samples were contaminated, whereas 35% of valve-culture samples were contaminated. PCR is more sensitive and specific than valve culture, and a valuable supplement to the existing analyses of valve tissue. PCR is necessary to identify the full spectrum of pathogens causing IE. In contrast to sensitivity of culture, sensitivity of PCR was independent of length of antibiotic treatment before surgery.

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“…Primers for the amplification have been published previously (19). All primers were manufactured by MWG Biotech, Germany.…”

Section: Methodsmentioning

confidence: 99%

Broad‐range PCR and sequencing in routine diagnosis of infective endocarditis

Voldstedlund¹,

Pedersen²,

Baandrup³

et al. 2008

APMIS

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“…In three patients (13)(14)(15), the disease was classified as aseptic spondylitis, as one component of an autoimmune disorder of unknown origin. 29 In the five remaining patients (16)(17)(18)(19)(20), spinal tumours were diagnosed by histology.…”

Section: Spinal Specimensmentioning

confidence: 99%

“…15 One case report has described identification of Fusobacterium species in a brain abscess, and another, Streptococcus pneumoniae in a subdural empyema, by the broad range bacterial PCR approach alone. 16 17 We have previously used broad range bacterial rDNA PCR to analyse 536 clinical samples of various tissues from patients admitted to hospital during the years 1994 to 1997. That work also included samples obtained during neurosurgery, but the patients involved were not further described nor the results specifically discussed.…”

mentioning

confidence: 99%

Aetiological diagnosis of brain abscesses and spinal infections: application of broad range bacterial polymerase chain reaction analysis

Kupila

Rantakokko‐Jalava²,

Jalava³

et al. 2003

Journal of Neurology, Neurosurgery &Amp; Psychiatry

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Objective: To evaluate the usefulness of the broad range bacterial rDNA polymerase chain reaction (PCR) method combined with DNA sequencing in the aetiological diagnosis of intracranial or spinal infections in neurosurgical patients. Methods: In addition to conventional methods, the broad range bacterial PCR approach was applied to examine pus or tissue specimens from cerebral or spinal lesions in patients treated in a neurosurgical unit for a clinical or neuroradiological suspicion of bacterial brain abscess or spondylitis. Results: Among the 44 patients with intracranial or spinal lesions, the final diagnosis suggested bacterial disease in 25 patients, among whom the aetiological agent was identified in 17. A causative bacterial species was identified only by the rDNA PCR method in six cases, by both the PCR methodology and bacterial culture in six cases, and by bacterial culture alone in five. All samples in which a bacterial aetiology was identified only by the PCR approach were taken during antimicrobial treatment, and in three patients the method yielded the diagnosis even after > 12 days of parenteral treatment. One case also identified by the PCR approach alone involved a brain abscess caused by Mycoplasma hominis, which is not readily cultured by routine methods. Conclusions: In patients with brain abscesses and spinal infections, the broad range bacterial rDNA PCR approach may be the only method to provide an aetiological diagnosis when the patient is receiving antimicrobial treatment, or when the causative agent is fastidious.

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“…Es handelt sich dabei einmalwie bei unserem Patienten -um die Detektion von Fusobacterium spp. [7], im zweiten Fall wurde Streptococcus pneumoniae bestimmt [6], während die Kulturen in beiden Fällen kein Bakterienwachstum zeigten. Beide Patienten standen unter einer empirischen Antibiotikatherapie.…”

Section: Diskussionunclassified

Erregerdetektion und -identifikation bei einem Hirnabszess durch molekulargenetische Methoden

et al. 2004

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Brain abscesses are life-threatening and detection and identification of the causative pathogens are crucial for substantiating the diagnosis and for selecting the optimal antibiotic regimen. In approximately 20% of the patients microbiological cultures of abscess material remain sterile. The polymerase chain reaction (PCR) provides a methodological alternative, but data about the use of broad spectrum PCR assays to detect the causative pathogens in brain abscesses are rare. We report on the case of a 65-years-old patient with a brain abscess caused by Fusobacterium spp., which was only diagnosed by broad spectrum PCR. To our knowledge this is the second report about a brain abscess, where Fusobacterium spp. was identified only by broad spectrum PCR and subsequent DNA sequencing.

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Eubacterial approach to the diagnosis of bacterial infection

Cited by 8 publications

References 7 publications

Broad‐range PCR and sequencing in routine diagnosis of infective endocarditis

Broad‐range PCR and sequencing in routine diagnosis of infective endocarditis

Aetiological diagnosis of brain abscesses and spinal infections: application of broad range bacterial polymerase chain reaction analysis

Erregerdetektion und -identifikation bei einem Hirnabszess durch molekulargenetische Methoden

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