Sensitivity and specificity of nested and real-time PCR for the detection of Pneumocystis jiroveci in clinical specimens

Álvarez-Martínez, Míriam J.; Miró, José M.; Valls, María Eugenia; Moreno, Asunción; Rivas, Paula; Solé, Manel; Benito, Natividad; Domingo, Peré; Muñoz, Carmen; Rivera, E.; Zar, Heather J.; Wissmann, Gustavo; Diehl, Ada Rs; Prolla, João Carlos; Anta, M. T. Jiménez de; Gatell, José M.; Wilson, Philip; Meshnick, Steven R.

doi:10.1016/j.diagmicrobio.2006.03.010

Cited by 97 publications

(58 citation statements)

References 32 publications

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“…Contrary to the findings of some studies, no patient with a negative qPCR result was observed among patients with PCP in the area of the Nice University Hospital between April 2008 and October 2013 (39)(40)(41)(42)(43)(44). All patients with positive microscopy results were diagnosed with PCP and had C T values below the threshold for exclusion of a diagnosis of PCP.…”

Section: Discussioncontrasting

confidence: 49%

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Detection of Pneumocystis jirovecii by Quantitative PCR To Differentiate Colonization and Pneumonia in Immunocompromised HIV-Positive and HIV-Negative Patients

et al. 2016

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dPneumocystis jirovecii pneumonia (PCP) is an acute and life-threatening lung disease caused by the fungus Pneumocystis jirovecii. The presentation of PCP in HIV-positive patients is well-known and consists of a triad of dyspnea, fever, and cough, whereas the presentation of PCP in HIV-negative patients is atypical and consists of a sudden outbreak, O 2 desaturation, and a rapid lethal outcome without therapy. Despite the availability of direct and indirect identification methods, the diagnosis of PCP remains difficult. The cycle threshold (C T ) values obtained by quantitative PCR (qPCR) allow estimation of the fungal burden. The more elevated that the fungal burden is, the higher the probability that the diagnosis is pneumonia. The purposes of the present study were to evaluate the C T values to differentiate colonization and pneumonia in a population of immunocompromised patients overall and patients stratified on the basis of their HIV infection status. Testing of bronchoalveolar lavage (BAL) fluid samples from the whole population of qPCR-positive patients showed a mean C T value for patients with PCP of 28 (95% confidence interval [CI], 26 to 30) and a mean C T value for colonized patients of 35 (95% CI, 34 to 36) (P < 10 ؊3 ). For the subgroup of HIV-positive patients, we demonstrated that a C T value below 27 excluded colonization and a C T value above 30 excluded PCP with a specificity of 100% and a sensitivity of 80%, respectively. In the subgroup of HIV-negative patients, we demonstrated that a C T value below 31 excluded colonization and a C T value above 35 excluded PCP with a specificity of 80% and a sensitivity of 80%, respectively. Thus, qPCR of BAL fluid samples is an important tool for the differentiation of colonization and pneumonia in P. jirovecii-infected immunocompromised patients and patients stratified on the basis of HIV infection status with different C T values.

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Section: Discussioncontrasting

confidence: 49%

“…The more elevated that the fungal burden is, the higher the probability that the diagnosis is pneumonia (37,38). Thus, the use of qPCR could greatly increase the specificity of detection with a good sensitivity even if it could not detect all cases of PCP (39)(40)(41)(42)(43)(44).…”

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confidence: 99%

Detection of Pneumocystis jirovecii by Quantitative PCR To Differentiate Colonization and Pneumonia in Immunocompromised HIV-Positive and HIV-Negative Patients

et al. 2016

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“…Indeed, Pneumocystis jirovecii cannot be cultured, and its detection is based on staining methods using respiratory specimens, which suffer from low sensitivity (214). Consequently, many studies have evaluated the performance of PCR on respiratory specimens for PCP diagnosis, and the results seem promising, with sensitivity values as high as 100% (91,(215)(216)(217)(218)(219)(220)(221)(222)(223)(224). However, in the case of PCP, colonization is a significant issue, with rates as high as 22% in high-risk populations (225), and more importantly, since DNA is fairly stable, it is difficult to distinguish between active and previous infections (226).…”

Section: Pcrmentioning

confidence: 99%

“…Studies have shown that the mean concentration of DNA from BAL fluid samples can be manifold higher in infected individuals than in asymptomatic carriers (219). Thus, many different real-time PCR assays have been developed and evaluated, with studies showing sensitivity values comparable to those of conventional PCR techniques and consistently higher than 80% (91,216,217,220,222,224). The real breakthrough, though, is found in the comparison of specificity values, with reports indicating a specificity of real-time PCR that can reach 98% and that is superior to those of conventional PCR techniques at cutoff points selected to not affect sensitivity (216).…”

Section: Pcrmentioning

confidence: 99%

Molecular and Nonmolecular Diagnostic Methods for Invasive Fungal Infections

et al. 2014

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SUMMARY Invasive fungal infections constitute a serious threat to an ever-growing population of immunocompromised individuals and other individuals at risk. Traditional diagnostic methods, such as histopathology and culture, which are still considered the gold standards, have low sensitivity, which underscores the need for the development of new means of detecting fungal infectious agents. Indeed, novel serologic and molecular techniques have been developed and are currently under clinical evaluation. Tests like the galactomannan antigen test for aspergillosis and the β-glucan test for invasive Candida spp. and molds, as well as other antigen and antibody tests, for Cryptococcus spp., Pneumocystis spp., and dimorphic fungi, have already been established as important diagnostic approaches and are implemented in routine clinical practice. On the other hand, PCR and other molecular approaches, such as matrix-assisted laser desorption ionization (MALDI) and fluorescence in situ hybridization (FISH), have proved promising in clinical trials but still need to undergo standardization before their clinical use can become widespread. The purpose of this review is to highlight the different diagnostic approaches that are currently utilized or under development for invasive fungal infections and to identify their performance characteristics and the challenges associated with their use.

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“…This variability is specially related to the skills of the team inducing the sputum [76]. Several PCR assays have been developed for the diagnosis of PCP, and have been tested on BAL, induced sputum and noninvasive oral wash specimens [77]. In general, PCR assays have been more sensitive but less specific for diagnosis of PCP than traditional microscopic methods.…”

Section: Bacterial Pneumoniamentioning

confidence: 99%

Pulmonary infections in HIV-infected patients: an update in the 21st century

Benito¹,

Moreno²,

Miró³

et al. 2011

Eur Respir J

176

216

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From the first descriptions of HIV/AIDS, the lung has been the site most frequently affected by the disease. Most patients develop a pulmonary complication during the history of HIV infection, mainly of infectious aetiology.Important changes in the epidemiology of HIV-related pulmonary infections have occurred. Overall, prescription of Pneumocystis jirovecii prophylaxis and the introduction of highly active antiretroviral therapy (HAART) are the main causes.Currently, the most frequent diagnosis in developed countries is bacterial pneumonia, especially pneumococcal pneumonia, the second most frequent cause is Pneumocystis pneumonia and the third is tuberculosis. However, in Africa, tuberculosis could be the most common pulmonary complication of HIV.Pulmonary infections remain one of the most important causes of morbidity and mortality in these patients, and the first cause of hospital admission in the HAART era. Achieving an aetiological diagnosis of pulmonary infection in these patients is important due to its prognostic consequences.

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Sensitivity and specificity of nested and real-time PCR for the detection of Pneumocystis jiroveci in clinical specimens

Cited by 97 publications

References 32 publications

Detection of Pneumocystis jirovecii by Quantitative PCR To Differentiate Colonization and Pneumonia in Immunocompromised HIV-Positive and HIV-Negative Patients

Detection of Pneumocystis jirovecii by Quantitative PCR To Differentiate Colonization and Pneumonia in Immunocompromised HIV-Positive and HIV-Negative Patients

Molecular and Nonmolecular Diagnostic Methods for Invasive Fungal Infections

Pulmonary infections in HIV-infected patients: an update in the 21st century

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