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Chun-Li An
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Abstract Background Pneumocystis jirovecii is an opportunistic pathogen fungus and the role of its colonization in respiratory diseases has become a popular focus in recent years. The aim of present study is to develop an improved loop-mediated isothermal amplification (LAMP) assay for detection of Pneumocystis jirovecii (P. jirovecii) DNA and use it to examine the prevalence and association of P. jirovecii colonization among non-HIV patients with various pulmonary diseases. Methods We modified the previously reported LAMP assay for P. jirovecii by adding real-time detection. This method was used to detect P. jirovecii colonization in respiratory samples collected from 403 non-HIV patients with various respiratory diseases enrolled from 5 hospitals in China. We determined the prevalence of P. jirovecii colonization in 7 types of pulmonary diseases and assessed the association of P. jirovecii colonization with clinical characteristics of these diseases. Results The new LAMP assay showed no cross-reactivity with other common respiratory microbes and was 1,000 times more sensitive than that of conventional PCR. Using the new LAMP assay, we detected P. jirovecii colonization in 281 (69.7%) of the 403 patients enrolled. P. jirovecii colonization was more common in interstitial lung diseases than in chronic obstructive pulmonary disease (COPD) (84.6% vs 64.5%, P < 0.05). Patients with acute exacerbation of COPD had a higher prevalence of P. jirovecii colonization compared to patients with stabilized COPD (67.4% vs 43.3%, P < 0.05). P. jirovecii colonization was associated with decreased pulmonary function, increased levels of 1,3-β-D-glucan and C-reactive protein, and decreased levels of CD4+ T-cell counts (P < 0.05 for each). Approximately 70% of P. jirovecii colonized patients had confections with other fungi or bacteria. Conclusions We developed an improved LAMP assay for detecting P. jirovecii. Our multi-center study of 403 patients supports that P. jirovecii colonization is a risk factor for the development of pulmonary diseases and highlights the need to further study the pathogenesis and transmission of P. jirovecii colonization in pulmonary diseases. Keywords: P. jirovecii; colonization; respiratory diseases; loop-mediated isothermal amplification.
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CURRENT STATUS: Published
View the published article at BMC Pulmonary Medicine.
No community comments so far
Editorial decision: Minor revision
On 10 Mar, 2020
Editor assigned
On 06 Mar, 2020
Submission checks complete
On 05 Mar, 2020
Editor invited
On 05 Mar, 2020
No comments provided
Editorial decision: Minor revision
On 02 Mar, 2020
Review #2 received
Received 24 Feb, 2020
Reviewer #2 agreed
On 13 Feb, 2020
Review #1 received
Received 09 Feb, 2020
Reviewer #1 agreed
On 15 Jan, 2020
Reviewers invited
Invitations sent on 10 Jan, 2020
Editor assigned
On 06 Jan, 2020
Submission checks complete
On 05 Jan, 2020
Editor invited
On 05 Jan, 2020
No comments provided
Editorial decision: Major revision
On 13 Dec, 2019
Review #2 received
Received 11 Dec, 2019
Review #1 received
Received 27 Nov, 2019
Reviewer #2 agreed
On 25 Nov, 2019
Reviewers invited
Invitations sent on 23 Nov, 2019
Reviewer #1 agreed
On 23 Nov, 2019
Editor assigned
On 07 Oct, 2019
Submission checks complete
On 06 Oct, 2019
Editor invited
On 06 Oct, 2019
Version 1
Posted 20 Jun, 2019
No comments provided
Review #2 received
Received 06 Sep, 2019
Editorial decision: Major revision
On 06 Sep, 2019
Review #1 received
Received 20 Aug, 2019
Reviewer #2 agreed
On 20 Jul, 2019
Reviewer #1 agreed
On 14 Jul, 2019
Reviewers invited
Invitations sent on 01 Jul, 2019
Submission checks complete
On 18 Jun, 2019
Editor invited
On 10 Jun, 2019
Editor assigned
On 10 Jun, 2019
First submitted
On 04 Jun, 2019
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Subject Areas
Pulmonology
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A new preprint design is coming!
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See the published version of this preprint at BMC Pulmonary Medicine.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research article
Chun-Li An
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
View the published article at BMC Pulmonary Medicine.
No community comments so far
Editorial decision: Minor revision
On 10 Mar, 2020
Editor assigned
On 06 Mar, 2020
Submission checks complete
On 05 Mar, 2020
Editor invited
On 05 Mar, 2020
No comments provided
Editorial decision: Minor revision
On 02 Mar, 2020
Review #2 received
Received 24 Feb, 2020
Reviewer #2 agreed
On 13 Feb, 2020
Review #1 received
Received 09 Feb, 2020
Reviewer #1 agreed
On 15 Jan, 2020
Reviewers invited
Invitations sent on 10 Jan, 2020
Editor assigned
On 06 Jan, 2020
Submission checks complete
On 05 Jan, 2020
Editor invited
On 05 Jan, 2020
No comments provided
Editorial decision: Major revision
On 13 Dec, 2019
Review #2 received
Received 11 Dec, 2019
Review #1 received
Received 27 Nov, 2019
Reviewer #2 agreed
On 25 Nov, 2019
Reviewers invited
Invitations sent on 23 Nov, 2019
Reviewer #1 agreed
On 23 Nov, 2019
Editor assigned
On 07 Oct, 2019
Submission checks complete
On 06 Oct, 2019
Editor invited
On 06 Oct, 2019
Version 1
Posted 20 Jun, 2019
No comments provided
Review #2 received
Received 06 Sep, 2019
Editorial decision: Major revision
On 06 Sep, 2019
Review #1 received
Received 20 Aug, 2019
Reviewer #2 agreed
On 20 Jul, 2019
Reviewer #1 agreed
On 14 Jul, 2019
Reviewers invited
Invitations sent on 01 Jul, 2019
Submission checks complete
On 18 Jun, 2019
Editor invited
On 10 Jun, 2019
Editor assigned
On 10 Jun, 2019
First submitted
On 04 Jun, 2019
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Pulmonology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at BMC Pulmonary Medicine.
Abstract Background Pneumocystis jirovecii is an opportunistic pathogen fungus and the role of its colonization in respiratory diseases has become a popular focus in recent years. The aim of present study is to develop an improved loop-mediated isothermal amplification (LAMP) assay for detection of Pneumocystis jirovecii (P. jirovecii) DNA and use it to examine the prevalence and association of P. jirovecii colonization among non-HIV patients with various pulmonary diseases. Methods We modified the previously reported LAMP assay for P. jirovecii by adding real-time detection. This method was used to detect P. jirovecii colonization in respiratory samples collected from 403 non-HIV patients with various respiratory diseases enrolled from 5 hospitals in China. We determined the prevalence of P. jirovecii colonization in 7 types of pulmonary diseases and assessed the association of P. jirovecii colonization with clinical characteristics of these diseases. Results The new LAMP assay showed no cross-reactivity with other common respiratory microbes and was 1,000 times more sensitive than that of conventional PCR. Using the new LAMP assay, we detected P. jirovecii colonization in 281 (69.7%) of the 403 patients enrolled. P. jirovecii colonization was more common in interstitial lung diseases than in chronic obstructive pulmonary disease (COPD) (84.6% vs 64.5%, P < 0.05). Patients with acute exacerbation of COPD had a higher prevalence of P. jirovecii colonization compared to patients with stabilized COPD (67.4% vs 43.3%, P < 0.05). P. jirovecii colonization was associated with decreased pulmonary function, increased levels of 1,3-β-D-glucan and C-reactive protein, and decreased levels of CD4+ T-cell counts (P < 0.05 for each). Approximately 70% of P. jirovecii colonized patients had confections with other fungi or bacteria. Conclusions We developed an improved LAMP assay for detecting P. jirovecii. Our multi-center study of 403 patients supports that P. jirovecii colonization is a risk factor for the development of pulmonary diseases and highlights the need to further study the pathogenesis and transmission of P. jirovecii colonization in pulmonary diseases. Keywords: P. jirovecii; colonization; respiratory diseases; loop-mediated isothermal amplification.
Figure 1
Figure 2
Figure 3
This is a list of supplementary files associated with this preprint. Click to download.
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