Detection of Mycoplasma pneumoniae by Polymerase Chain Reaction in Lung Aspirates from Patients with Community-Acquired Pneumonia

Falguera, Miquel; Nogués, A; Ruiz-González, Agustı́n; García, M.; Puig, Teresa

doi:10.1378/chest.110.4.972

Cited by 21 publications

(15 citation statements)

References 30 publications

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“…Development of molecular-based testing such as the PCR assay has lessened the importance of culture as a means for detecting M. pneumoniae. Studies since the late 1980s using simulated clinical specimens, animal models, and later clinical trials have validated the ability of PCR to detect M. pneumoniae, often in conjunction with serology and/or culture (43,46,47,53,96,108,111,127,180,199,211,215,239,337,382,417,443). The same types of clinical specimens that can undergo culture can also be tested by PCR.…”

Section: Microbiological Testsmentioning

confidence: 99%

“…However, in a case with a negative PCR assay and a positive culture (or serology), the presence of inhibitors or some other technical problem with the PCR assay must be considered (127,199,346). Reznikov et al (346) showed that PCR inhibition was much more likely to occur with nasopharyngeal aspirates than with throat swabs and recommended the latter specimen for diagnostic purposes for M. pneumoniae.…”

Section: Microbiological Testsmentioning

confidence: 99%

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Mycoplasma pneumoniaeand Its Role as a Human Pathogen

2004

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Mycoplasma pneumoniae is a unique bacterium that does not always receive the attention it merits considering the number of illnesses it causes and the degree of morbidity associated with it in both children and adults. Serious infections requiring hospitalization, while rare, occur in both adults and children and may involve multiple organ systems. The severity of disease appears to be related to the degree to which the host immune response reacts to the infection. Extrapulmonary complications involving all of the major organ systems can occur in association with M. pneumoniae infection as a result of direct invasion and/or autoimmune response. The extrapulmonary manifestations are sometimes of greater severity and clinical importance than the primary respiratory infection. Evidence for this organism's contributory role in chronic lung conditions such as asthma is accumulating. Effective management of M. pneumoniae infections can usually be achieved with macrolides, tetracyclines, or fluoroquinolones. As more is learned about the pathogenesis and immune response elicited by M. pneumoniae, improvement in methods for diagnosis and prevention of disease due to this organism may occur

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Section: Microbiological Testsmentioning

confidence: 99%

Section: Microbiological Testsmentioning

confidence: 99%

Mycoplasma pneumoniaeand Its Role as a Human Pathogen

2004

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“…Specimens suitable for the detection of M. pneumoniae include sputum (74) and bronchoalveolar lavage (BAL) specimens (40,66), nasopharyngeal and throat swabs (25,78), nasopharyngeal aspirates (20,28,36), tracheal aspirates (1,26), pleural fluid specimens (52), and transthoracic needle aspirations (17). More unusual specimens, such as nasal polyps (29), open-lung biopsies, and autopsy specimens (65), have also been tested.…”

Section: Technical Aspectsmentioning

confidence: 99%

“…Different extraction methods with and without sample pretreatment have been described: (i) dilution of the sample with 0.9% sodium chloride, followed by the addition of sodium dodecyl sulfate, extraction with phenolchloroform, and precipitation with ammonium acetate and ethanol (28); (ii) pretreatment with proteinase K (15), followed by phenol-chloroform or phenol-chloroform-isoamyl alcohol extraction and ethanol precipitation (10); (iii) Boom extraction (5) on protease-treated (47) or untreated samples (62); (iv) treatment with Sputazyme (Kobayashi Pharmaceutical Co., Tokyo, Japan), followed by proteinase K digestion (34); (v) incubation with Chelex (Bio-Rad Laboratories, Richmond, Calif.) and sodium azide (40); (vi) treatment by sonication or boiling (7); (vii) phenol-chloroform-isoamyl alcohol extraction, followed by ether extraction (79); and (viii) phenolchloroform extraction and precipitation by sodium acetate or sodium chloride (17).…”

Section: Technical Aspectsmentioning

confidence: 99%

Molecular Diagnosis of Mycoplasma pneumoniae Respiratory Tract Infections

et al. 2003

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2Mycoplasma pneumoniae is responsible for 10 to 20% of the cases of community-acquired pneumonia and has been associated with acute exacerbations of asthma (22). M. pneumoniae is also implicated in mild acute respiratory infections, such as sore throat, pharyngitis, rhinitis, and tracheobronchitis (2).Correct diagnosis of M. pneumoniae infections is important to allow the appropriate antibiotic treatment of patients, since it is impossible to identify a M. pneumoniae infection solely on the basis of clinical signs and symptoms. It should decrease inappropriate use of antibiotics, influence the patient outcome by reduction of morbidity and mortality, and improve our knowledge of the prevalence of the causes of so-called atypical pneumonia.Conventional assays for the detection of M. pneumoniae have their limitations, resulting in the need for more accurate diagnostic methods. Culture is time-consuming and relatively insensitive, because M. pneumoniae grows slowly in vitro, requiring 2 to 5 weeks for colonies to become visible. Serological methods, particularly the complement fixation (CF) test, are most widely used. The sensitivity of these assays depends on whether the first serum sample is collected early or late after the onset of disease and on the availability of paired serum samples collected with an interval of 2 to 3 weeks. Immunoglobulin M (IgM) assays which are more sensitive than the CF test have been developed, but the IgM response may be nonspecific (61) or absent, particularly in adults (70). Hybridization with DNA probes has also been proposed as a rapid and specific procedure to replace culture, but it lacks sensitivity (35).Nucleic acid amplification techniques (NAATs) have the potential to produce rapid, sensitive, and specific results, allowing early appropriate antibiotic therapy.In the absence of a reference method, the so-called "gold standard" for the diagnosis of an M. pneumoniae infection, either an expanded gold standard or the technique of latent class analysis (LCA) should be applied to calculate the sensitivity and specificity of the available diagnostic tests. The technique of LCA can be used if at least three independent techniques can be compared. Thus far, only a few PCR tests and a limited number of studies applying culture, serology, and several NAATs targeting different genes to detect M. pneumoniae have been adequately evaluated.Since NAATs targeting DNA can detect both viable and nonviable organisms, detecting RNA by reverse transcriptase PCR (RT-PCR) or nucleic acid sequence-based amplification (NASBA) may be a useful method to identify productive M. pneumoniae infections.The possible long-term carrier state of M. pneumoniae in the respiratory tract may hinder the evaluation of different diagnostic tests for the diagnosis of acute infections.An overview of the peer-reviewed literature on the use of NAATs to detect M. pneumoniae since 1989 is given. Search combinations were M. pneumoniae and PCR, M. pneumoniae and diagnosis, and M. pneumoniae and amplification. This minireview...

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“…For MP the sensitivity of serology is also low in the acute phase of the infection [28]. High antibody levels may persist for long periods [3,29]. Here, we used both PCR and antigen detection for diagnosis, because antibody detection did not prove reliable on account of the difficulty of obtaining paired sera.…”

Section: Discussionmentioning

confidence: 99%