Rapid detection of Staphylococcus aureus bacteremia and methicillin resistance by real-time PCR in whole blood samples

Wellinghausen, Nele; Siegel, Dunja; Gebert, Susanne; Winter, Juliane

doi:10.1007/s10096-009-0723-7

Cited by 30 publications

(16 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Notably, PCR has been reported to have difficulties with whole blood specimen protocols, despite its inherent promise [21]. Staphylococcal bacteremia has been estimated to have a lowend range of 1–30 CFU per ml of blood, this low concentration is sufficient for obtaining growth in blood culture bottles but below the sensitivity of a PCR assay.…”

Section: Resultsmentioning

confidence: 99%

PNA-based microbial pathogen identification and resistance marker detection

Smolina¹,

Miller²,

Frank‐Kamenetskii³

2010

Artificial DNA: PNA & XNA

View full text Add to dashboard Cite

Section: Resultsmentioning

confidence: 99%

PNA-based microbial pathogen identification and resistance marker detection

Smolina¹,

Miller²,

Frank‐Kamenetskii³

2010

Artificial DNA: PNA & XNA

View full text Add to dashboard Cite

“…Human blood cells are lysed and the released human DNA degraded, followed by a bindwash-elute procedure for the isolation of pathogen DNA. The (37).…”

Section: Discussionmentioning

confidence: 99%

Diagnosis of Bacteremia in Whole-Blood Samples by Use of a Commercial Universal 16S rRNA Gene-Based PCR and Sequence Analysis

et al. 2009

View full text Add to dashboard Cite

In a prospective, multicenter study of 342 blood samples from 187 patients with systemic inflammatory response syndrome, sepsis, or neutropenic fever, a new commercial PCR test (SepsiTest; Molzym) was evaluated for rapid diagnosis of bacteremia. The test comprises a universal PCR from the 16S rRNA gene, with subsequent identification of bacteria from positive samples by sequence analysis of amplicons. Compared to blood culture (BC), the diagnostic sensitivity and specificity of the PCR were 87.0 and 85.8%, respectively. Considering the 34 BC-positive patients, 28 were also PCR positive in at least one of the samples, resulting in a patient-related sensitivity of 82.4%. The concordance of PCR and BC for both positive and negative samples was (47 ؉ 247)/342, i.e., 86.0%. In total, 31 patients were PCR/sequencing positive and BC negative, in whom the PCR result was judged as possible or probable to true bacteremia in 25. In conclusion, the PCR approach facilitates the detection of bacteremia in blood samples within a few hours. Despite the indispensability of BC diagnostics, the rapid detection of bacteria by SepsiTest appears to be a valuable tool, allowing earlier pathogen-adapted antimicrobial therapy in critically ill patients.Bloodstream infection is a life-threatening condition with a high mortality rate, especially in intensive care and neutropenic patients (5,19,35,38). Pathogenic bacteria are the most frequent causes of bloodstream infection, although fungi can also be isolated in a minority of patients (7,17,21,32,34). Currently, inoculation of blood cultures (BC) is the standard method for microbiological diagnosis of bloodstream infections. However, the limitations of BC include relatively low sensitivities and a long time-to-result for detection and identification of the pathogen, generally over 2 days, and even longer for fastidious organisms (13,20,27).In contrast, DNA-based procedures may offer faster and more reliable diagnoses (3, 30). PCR amplification of microbial genes, followed by detection of amplified products by gel electrophoresis or real-time PCR monitoring using fluorescent dyes or target-directed fluorescent probes, is a quick process allowing pathogen detection within a few hours (18). Identification of microorganisms can be performed by PCR algorithms, taxon-specific oligonucleotide microarrays, or sequencing amplicons (30).PCR amplification of conserved regions of the bacterial genome, in particular the 16S rRNA gene, combined with sequence analysis is a well-established technique for the identification of bacterial pathogens (18). The main advantages of targeting the 16S rRNA gene are the broad range of pathogens detectable and the independence of this method from the in vitro viability of strains (6). The high sensitivity of detection by PCR of bacterial DNA (15) suggests its use in the diagnosis of bacteremia (16). Initial disadvantages of PCR, notably the incidence of false-positive results from bacterial DNA contaminating PCR reagents (4, 39), have been counteracted by the devel...

show abstract

“…The development and implementation of molecular diagnostic tests for pneumonia has been a major advance in the microbiological diagnosis of respiratory pathogens in the last ten years [85,86,87,88]. Molecular tests help us identify a specific pathogen or help distinguish between bacterial and viral infection and provide information about antibiotic susceptibility patterns, monitor the response to antibiotic therapy, assess prognosis, aid antimicrobial stewardship, and give information for disease surveillance.…”

Section: Laboratory Diagnosis Of Pneumoniamentioning

confidence: 99%

Microbial Etiology of Pneumonia: Epidemiology, Diagnosis and Resistance Patterns

Cillóniz

Martín‐Loeches

García-Vidal

et al. 2016

IJMS

219

161

View full text Add to dashboard Cite

Globally, pneumonia is a serious public health concern and a major cause of mortality and morbidity. Despite advances in antimicrobial therapies, microbiological diagnostic tests and prevention measures, pneumonia remains the main cause of death from infectious disease in the world. An important reason for the increased global mortality is the impact of pneumonia on chronic diseases, along with the increasing age of the population and the virulence factors of the causative microorganism. The increasing number of multidrug-resistant bacteria, difficult-to-treat microorganisms, and the emergence of new pathogens are a major problem for clinicians when deciding antimicrobial therapy. A key factor for managing and effectively guiding appropriate antimicrobial therapy is an understanding of the role of the different causative microorganisms in the etiology of pneumonia, since it has been shown that the adequacy of initial antimicrobial therapy is a key factor for prognosis in pneumonia. Furthermore, broad-spectrum antibiotic therapies are sometimes given until microbiological results are available and de-escalation cannot be performed quickly. This review provides an overview of microbial etiology, resistance patterns, epidemiology and microbial diagnosis of pneumonia.

show abstract

Rapid detection of Staphylococcus aureus bacteremia and methicillin resistance by real-time PCR in whole blood samples

Cited by 30 publications

References 10 publications

PNA-based microbial pathogen identification and resistance marker detection

PNA-based microbial pathogen identification and resistance marker detection

Diagnosis of Bacteremia in Whole-Blood Samples by Use of a Commercial Universal 16S rRNA Gene-Based PCR and Sequence Analysis

Microbial Etiology of Pneumonia: Epidemiology, Diagnosis and Resistance Patterns

Contact Info

Product

Resources

About