Evaluation of a multiplex PCR for bacterial pathogens applied to bronchoalveolar lavage

Strålin, Kristoffer; Korsgaard, J.; Olcén, Per

doi:10.1183/09031936.06.00006106

Cited by 56 publications

(43 citation statements)

References 38 publications

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“…This high detection rate through PCR is similar to the study in which respiratory microorganisms were detected in 97% of the children with CAP by Honkinen et al [12]. Strålin et al suggested that bronchoalveolar lavage multiplex PCR for S. pneumonia, M. pneumoniae and C. pneumoniae appears to be a useful etiological tool in lower respiratory tract infection patients and this method may be a valuable supplement to bronchoalveolar lavage culture [11]. Another probable reason for high etiologic agent recovery may be the severity of disease and the poor immune response in the elder patients requiring hospitalization.…”

Section: Discussionsupporting

confidence: 69%

See 1 more Smart Citation

Bacterial and viral etiology in hospitalized community acquired pneumonia with molecular methods and clinical evaluation

Serin

Pullukçu

Çiçek

et al. 2014

J Infect Dev Ctries

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Introduction: Polymerase chain reaction (PCR) method has improved the diagnosis rates for patients with community-acquired pneumonia (CAP). We aimed to evaluate the bacterial and viral etiology of hospitalized CAP cases and compare clinical and laboratory findings of patients with pure bacterial and bacterial and viral (mixed) infections. Methodology: A total of 55 patients hospitalized with CAP were enrolled into the prospective study between February 2010 and December 2010. Clinical and laboratory follow-up were performed on days 0, 7 and 14. Deep tracheal aspiration samples were examined for bacterial and viral pathogens by multiplex PCR, and standard bacteriological culture method. Results: The etiological identification rate in 50 patients for bacteria, viruses and mixed virus-bacteria combination by PCR were 62%, 4%, 32%, respectively and 60% in 55 patients by bacterial culture method. Streptococcus pneumoniae concomitant with Haemophilus influenzae (36%) and rhinovirus (16%) was very common, whereas atypical pathogens (only Mycoplasma pneumoniae) were rare (6%). Rhinovirus was the most common viral agent (20%). Recently identified viruses, human coronavirus HKU1 and human bocavirus were not detected except for human metapneumovirus (one case). There was no significant difference in terms of mean age, immune status, leukocyte count, Creactive protein (CRP) values, hospitalization duration and CURB-65 score between bacterial and mixed viral-bacterial detections. Advanced age (p < 0.01) and higher CURB-65 score (p = 0.01) were found to be associated with increased mortality. Conclusion: Concomitance of bacterial and viral agents is frequent and resemble with bacterial infections alone. Further studies are needed for the clinical significance of mixed detections.

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Section: Discussionsupporting

confidence: 69%

“…This is probably due to i) molecular tests which provided doubled increase in identification rate [6,11] ii) respiratory samples obtained by protected deep tracheal aspiration. This high detection rate through PCR is similar to the study in which respiratory microorganisms were detected in 97% of the children with CAP by Honkinen et al [12].…”

Section: Discussionmentioning

confidence: 99%

Bacterial and viral etiology in hospitalized community acquired pneumonia with molecular methods and clinical evaluation

Serin

Pullukçu

Çiçek

et al. 2014

J Infect Dev Ctries

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“…However, the study specimens showed virtually no carriage of the other nine bacterial and viral pathogens. The findings of this work, combined with those in previous reports showing good sensitivities and specificities for symptomatic patients (11,20,24,25,29,40), suggest that molecular assays testing for these nine pathogens in respiratory secretions will have good utility in clinical practice. The low rate of detection of NA in serum is reassuring in the presence of comparably high rates of detection in specimens from the nasopharynx and skin, sites that have been implicated previously as potentially contributing to the detection of pathogens in serum.…”

Section: Discussionmentioning

confidence: 59%

Detection of 11 Common Viral and Bacterial Pathogens Causing Community-Acquired Pneumonia or Sepsis in Asymptomatic Patients by Using a Multiplex Reverse Transcription-PCR Assay with Manual (Enzyme Hybridization) or Automated (Electronic Microarray) Detection

et al. 2008

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Community-acquired pneumonia (CAP) and sepsis are important causes of morbidity and mortality. We describe the development of two molecular assays for the detection of 11 common viral and bacterial agents of CAP and sepsis: influenza virus A, influenza virus B, respiratory syncytial virus A (RSV A), RSV B, Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, Legionella micdadei, Bordetella pertussis, Staphylococcus aureus, and Streptococcus pneumoniae. Further, we report the prevalence of carriage of these pathogens in respiratory, skin, and serum specimens from 243 asymptomatic children and adults. The detection of pathogens was done using both a manual enzyme hybridization assay and an automated electronic microarray following reverse transcription and PCR amplification. The analytical sensitivities ranged between 0.01 and 100 50% tissue culture infective doses, cells, or CFU per ml for both detection methods. Analytical specificity testing demonstrated no significant cross-reactivity among 19 other common respiratory organisms. One hundred spiked "surrogate" clinical specimens were all correctly identified with 100% specificity (95% confidence interval, 100%). Overall, 28 (21.7%) of 129 nasopharyngeal specimens, 11 of 100 skin specimens, and 2 of 100 serum specimens from asymptomatic subjects tested positive for one or more pathogens, with S. pneumoniae and S. aureus giving 89% of the positive results. Our data suggest that asymptomatic carriage makes the use of molecular assays problematic for the detection of S. pneumoniae or S. aureus in upper respiratory tract secretions; however, the specimens tested showed virtually no carriage of the other nine viral and bacterial pathogens, and the detection of these pathogens should not be a significant diagnostic problem. In addition, slightly less sensitive molecular assays may have better correlation with clinical disease in the case of CAP.

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“…Over the past decade, several studies aimed to define parameters that allow the distinction between bacterial and viral infections causing LRTI. The methods used included polymerase chain reaction (PCR), procalcitonin determination and other clinical information [8,9,10,11,12,13,14,15]. However, the information obtained by medical history, physical examination, routine laboratory parameters and chest radiography is not sufficient for a clear differential diagnosis of the cause of LRTI.…”

Section: Introductionmentioning

confidence: 99%

Single and Multiple Viral Infections in Lower Respiratory Tract Infection

Gencay¹,

Roth²,

Müeller

et al. 2010

Respiration

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Background: Lower respiratory tract infection (LRTI) often leads to hospitalization, and it was indicated that causative viral infections are underestimated. Objectives: It was our aim to compare the frequency of 8 relevant viruses in 109 hospitalized LRTI patients and 144 healthy controls. Methods: Virus infection was determined by seroconversion and ELISA for anti-virus antibodies in repeated serum samples. Bacterial infection was diagnosed in respiratory specimens, blood cultures and urine. Results: The LRTI patient cohort consisted of 49 patients with community-acquired pneumonia, 30 patients with acute bronchitis and 30 chronic obstructive pulmonary disease patients with acute exacerbation. Viral infection was detected in 89 (82%) LRTI patients compared with 32 (22%) in healthy controls (relative risk 3.42, 95% confidence interval 2.48–4.72; p < 0.0001). The most frequent viral pathogens were: influenza B (23%), adenovirus (16%) and parainfluenza virus 3 (12%). Importantly, infections with more than 1 virus were detected in 63% (n = 57) of LRTI patients with viral infection, which represents 52% of all LRTI patients. No multiple virus infection was detected in the healthy controls. Patients with community-acquired pneumonia were more often infected with adenovirus and respiratory syncytial virus as compared with the other LRTI patients (p = 0.046 and 0.0009, respectively). Conclusions: There is a high incidence of single and multiple viral infections in LRTI patients requiring hospitalization. The data indicate the need for regular virus diagnosis and the development of point of care tools that enables a fast diagnosis of the most common viruses and bacteria. The data also imply the need to consider antiviral therapy in positive LRTI cases.

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Evaluation of a multiplex PCR for bacterial pathogens applied to bronchoalveolar lavage

Cited by 56 publications

References 38 publications

Bacterial and viral etiology in hospitalized community acquired pneumonia with molecular methods and clinical evaluation

Bacterial and viral etiology in hospitalized community acquired pneumonia with molecular methods and clinical evaluation

Detection of 11 Common Viral and Bacterial Pathogens Causing Community-Acquired Pneumonia or Sepsis in Asymptomatic Patients by Using a Multiplex Reverse Transcription-PCR Assay with Manual (Enzyme Hybridization) or Automated (Electronic Microarray) Detection

Single and Multiple Viral Infections in Lower Respiratory Tract Infection

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