scholarly journals Detection of Opportunistic Fungus Pneumocystis jirovecii Major Surface Glycoprotein (MSG) gene in HIV-AIDS Patients with Pneumoniae in Jakarta

2020 ◽  
Author(s):  
Conny Riana Tjampakasari ◽  
Andi Yasmon ◽  
Agus Sjahrurachman ◽  
Samsuridjal Djauzi
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Opportunistic Infections ◽  
Surface Glycoprotein ◽  
Pneumocystis Jirovecii ◽  
Molecular Diagnostic ◽  
Major Surface Glycoprotein ◽  
Major Surface ◽  
Suspected Pneumonia ◽  
Hiv Aids

Pneumocystis jirovecii is known to cause opportunistic infections in the lower respiratory tract in individuals with low immune systems, especially patient with HIV infection. The prevalence of P. jirovecii pneumonia (PjP) in various countries show varying numbers. In Indonesia, HIV cases continue to rise. However, the data in Indonesia concerning the case of PjP is very limited. Until now the prevalence of PjP in Indonesia is only based on clinical symptoms of the patient. Currently, diagnosis of PjP relies on microscopic examination. The disadvantage of this examination is not easy to do and has a high negative predictive value. Thus, this study was conducted to develop a molecular test to diagnose PjP infection in HIV-AIDS suspected pneumonia. Molecular diagnostic test aimed for Major Surface Glycoprotein (MSG) gene of P. jirovecii detection was done through real-time PCR against 100 sputum samples. Demographic data show that the prevalence of PjP infection in HIV-AIDS suspected pneumonia patients in Jakarta is 20.0%, male 75% within 31-40 y.o (35%), dominant (80%) from patients with CD4+ T-lymphocytes of 200-349 cells/µL. Molecular real-time PCR methods were shown to give five times sensitivity higher than Giemsa stain.   Keywords: P. jirovecii, HIV, real-time PCR

scholarly journals The Fungal PCR Initiative's evaluation of in-house and commercial Pneumocystis jirovecii qPCR assays: Toward a standard for a diagnostics assay

2019 ◽  
Vol 58 (6) ◽  
pp. 779-788 ◽  
Author(s):  
Maud Gits-Muselli ◽  
P Lewis White ◽  
Carlo Mengoli ◽  
Sharon Chen ◽  
Brendan Crowley ◽  
...  
Keyword(s):  
Target Genes ◽  
Clinical Care ◽  
Large Subunit ◽  
Reference Method ◽  
Pneumocystis Pneumonia ◽  
Surface Glycoprotein ◽  
Pneumocystis Jirovecii ◽  
Tubulin Genes ◽  
Major Surface Glycoprotein ◽  
Major Surface

Abstract Quantitative real-time PCR (qPCR) is increasingly used to detect Pneumocystis jirovecii for the diagnosis of Pneumocystis pneumonia (PCP), but there are differences in the nucleic acids targeted, DNA only versus whole nucleic acid (WNA), and also the target genes for amplification. Through the Fungal PCR Initiative, a working group of the International Society for Human and Animal Mycology, a multicenter and monocenter evaluation of PCP qPCR assays was performed. For the multicenter study, 16 reference laboratories from eight different countries, performing 20 assays analyzed a panel consisting of two negative and three PCP positive samples. Aliquots were prepared by pooling residual material from 20 negative or positive- P. jirovecii bronchoalveolar lavage fluids (BALFs). The positive pool was diluted to obtain three concentrations (pure 1:1; 1:100; and 1:1000 to mimic high, medium, and low fungal loads, respectively). The monocenter study compared five in-house and five commercial qPCR assays testing 19 individual BALFs on the same amplification platform. Across both evaluations and for all fungal loads, targeting WNA and the mitochondrial small sub-unit (mtSSU) provided the earliest Cq values, compared to only targeting DNA and the mitochondrial large subunit, the major surface glycoprotein or the beta-tubulin genes. Thus, reverse transcriptase-qPCR targeting the mtSSU gene could serve as a basis for standardizing the P. jirovecii load, which is essential if qPCR is to be incorporated into clinical care pathways as the reference method, accepting that additional parameters such as amplification platforms still need evaluation.

scholarly journals Use of Oropharyngeal Washes to Diagnose and Genotype Pneumocystis jirovecii

2015 ◽  
Vol 2 (3) ◽  
Author(s):  
Jonathan J. Juliano ◽  
Eric Barnett ◽  
Christian M. Parobek ◽  
Steve M. Taylor ◽  
Steven R. Meshnick ◽  
...  
Keyword(s):  
Clinical Symptoms ◽  
High Sensitivity ◽  
Diagnostic Techniques ◽  
Surface Glycoprotein ◽  
Pneumocystis Jirovecii ◽  
Major Surface Glycoprotein ◽  
Immunodeficiency Virus ◽  
Noninvasive Samples ◽  
Strain Type ◽  
Major Surface

Abstract Pneumocystis jirovecii is a symbiotic respiratory fungus that presents in 2 clinical forms: pneumonia in immunocompromised patients or colonization, defined by the presence of the organism without associated clinical symptoms. Currently, diagnosis requires invasive bronchoscopy, which may not be available in some settings and is inappropriate for detecting colonization in healthy individuals. Noninvasive diagnostic techniques and molecular strain typing tools that can be used on these samples are critical for conducting studies to better understand transmission. We evaluated 2 real-time polymerase chain reaction (PCR) assays targeting dihydropteroate synthase and the major surface glycoprotein for detection in 77 oropharyngeal washes (OPWs) from 43 symptomatic human immunodeficiency virus-infected patients who underwent bronchoscopy. We also evaluated the ability of a new microsatellite (MS) genotyping panel to strain type infections from these samples. Each PCR used individually provided a high sensitivity (>80%) for detection of pneumonia but a modest specificity (<70%). When used in combination, specificity was increased to 100% with a drop in sensitivity (74%). Concentration of organisms by PCR in the OPW tended to be lower in colonized individuals compared with those with pneumonia, but differences in concentration could not clearly define colonization in symptomatic individuals. Oropharyngeal wash samples were genotyped using 6 MSs with ≥4 alleles successfully genotyped in the majority of colonized patients and ≥5 alleles in patients with pneumonia. The MS profile was consistent over time within patients with serial OPWs analyzed. Microsatellite genotyping on noninvasive samples may aid in studying the molecular epidemiology of this pathogen without requiring invasive diagnostic techniques.

scholarly journals Serologic Responses to Recombinant Pneumocystis jirovecii Major Surface Glycoprotein among Ugandan Patients with Respiratory Symptoms

PLoS ONE ◽  
2012 ◽  
Vol 7 (12) ◽  
pp. e51545 ◽  
Author(s):  
Robert J. Blount ◽  
Leah G. Jarlsberg ◽  
Kieran R. Daly ◽  
William Worodria ◽  
J. Lucian Davis ◽  
...  
Keyword(s):  
Respiratory Symptoms ◽  
Surface Glycoprotein ◽  
Pneumocystis Jirovecii ◽  
Major Surface Glycoprotein ◽  
Major Surface

scholarly journals Analisis filogenetik gen mitochondrial large subunit (mtLSU) Pneumocystis jirovecii pada Pasien Human Immunodeficiency Virus (HIV) Terduga Pneumoniae di Jakarta

2020 ◽  
Vol 9 (1) ◽  
pp. 9-18
Author(s):  
Conny Riana Tjampakasari ◽  
Andi Yasmon ◽  
Agus Sjahrurachman ◽  
Samsuridjal Djauzi
Keyword(s):  
Human Immunodeficiency Virus ◽  
Hot Spot ◽  
Large Subunit ◽  
Surface Glycoprotein ◽  
Pneumocystis Jirovecii ◽  
Rt Pcr ◽  
Major Surface Glycoprotein ◽  
Immunodeficiency Virus ◽  
Polymerase Chain ◽  
Major Surface

Abstrak Pneumocystis jirovecii (P. jirovecii) merupakan patogen oportunistik yang penting pada pasien dengan gangguan kekebalan menurun khususnya human immunodeficiency virus (HIV). P. jirovecii tersebar dimanamana, menyebar melalui udara, dan menyerang sistem pernapasan atas. P. jirovecii mempunyai beberapa faktor virulensi, antara lain major surface glycoprotein (MSG) yang merupakan antigen yang paling banyak ditemukan di permukaan. Pendekatan biologi molekuler digunakan untuk mempelajari patogen ini karena hingga saat ini kultur belum dapat dilakukan. Penelitian ini bertujuan untuk memperoleh data genotip yang dapat dimanfaatkan sebagai dasar data demografi dan epidemiologi molekuler P. jirovecii di Indonesia. Dua puluh sampel sputum positif P. jirovecii pada real-time polymerase chain reaction (RT-PCR) dilakukan karakterisasi terhadap gen mitochondrial large subunit (mtLSU). Virulensi daerah hot spot gen mtLSU dianalisis dengan metode PCR dan sekuensing deoxyribonucleic acid (DNA). Diperoleh 30 strain dengan 7 varian didominasi oleh varian 3 yang bersirkulasi di Jakarta. Analisis filogenetik dengan strain negara lain menunjukkan strain Jakarta berkerabat dekat dengan strain Iran, India dan Korea. Kata kunci : Pneumocystis jirovecii, mtLSU, PCR, filogenetik Abstract Pneumocystis jirovecii (P. jirovecii) is an important opportunistic pathogen in immunocompromised patients especially human immunodeficiency virus (HIV). P. jirovecii is spread everywhere, spread through the air, and attacking the upper respiratory system. P. jirovecii has several virulence factors, including major surface glycoprotein (MSG) which is the most widely found on the surface antigen. The molecular biology approach is used to study this pathogen because until now culture cannot be done. This study aims to obtain genotype data that can be used as a basis for demographic and molecular epidemiological data of P. jirovecii in Indonesia. Twenty P. jirovecii positive sputum samples on real-time polymerase chain reaction (RT-PCR) were characterized by the mitochondrial large subunit (mtLSU) gene. Virulence of the mtLSU gene hot spot region was analyzed by PCR method and deoxyribonucleic acid (DNA) sequencing. Obtained 30 strains with 7 variants dominated by variant 3 circulating in Jakarta. Phylogenetic analyzed with strains of other countries shows that Jakarta strains are closely related to strains of Iran, India dan Korea. Keywords: Pneumocystis jirovecii, mtLSU, PCR, phylogenetic

scholarly journals Current State of Carbohydrate Recognition and C-Type Lectin Receptors in Pneumocystis Innate Immunity

2021 ◽  
Vol 12 ◽  
Author(s):  
Theodore J. Kottom ◽  
Eva M. Carmona ◽  
Andrew H. Limper
Keyword(s):  
Immune Response ◽  
Fungal Pathogens ◽  
Surface Glycoprotein ◽  
Pneumocystis Jirovecii ◽  
Pneumocystis Jirovecii Pneumonia ◽  
Lectin Receptors ◽  
Major Surface Glycoprotein ◽  
Current State ◽  
Major Surface ◽  
Wall Components

Pneumocystis jirovecii is one of the most common fungal pathogens in immunocompromised individuals. Pneumocystis jirovecii pneumonia (PJP) causes a significant host immune response that is driven greatly by the organism’s cell wall components including β-glucans and major surface glycoprotein (Msg). These ligands interact with a number of C-type lectin receptors (CLRs) leading to downstream activation of proinflammatory signaling pathways. This minireview provides a brief overview summarizing known CLR/Pneumocystis interactions.

scholarly journals Serum Antibody Levels to the Pneumocystis jirovecii Major Surface Glycoprotein in the Diagnosis of P. jirovecii Pneumonia in HIV+ Patients

PLoS ONE ◽  
2010 ◽  
Vol 5 (12) ◽  
pp. e14259 ◽  
Author(s):  
Kpandja Djawe ◽  
Laurence Huang ◽  
Kieran R. Daly ◽  
Linda Levin ◽  
Judy Koch ◽  
...  
Keyword(s):  
Serum Antibody ◽  
Surface Glycoprotein ◽  
Pneumocystis Jirovecii ◽  
Hiv Patients ◽  
Major Surface Glycoprotein ◽  
Major Surface ◽  
Antibody Levels
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