DETEKSI GEN MSG UNTUK PENINGKATKAN KETEPATAN DIAGNOSIS PNEUMOCYSTIS JIROVECII PADA PASIEN HIV DENGAN PNEUMONIA

Conny Riana Tjampakasari, Andi Yasmon, Fitrahwati Sudarmo

Abstract


Pneumocystis jirovecii is the cause of opportunistic infections in the lower respiratory tract of immunocompromised patients, especiallyin human immunodeficiencyvirus (HIV). Until now the case of P. jirovecii in Indonesia is not known with certainty because the data obtained only based on patient’s clinical condition. Culture is still on going research while microscopic as a gold standard has some limitation, among others, takes a long time in the process, requires special kills to work and interpret results. The diagnosis of P.jirovecii infection in Indonesia is still based on clinical and microscopic examination, which takes a long time, is less sensitive and specific. Based on this reason then developed rPCR that  more sensitive and specific. In addtionthe results obtained are less sensitive and spesific. Based on that, in this study developed real-time PCR (rPCR) molecular test that more sensitive and apesific using gene MajorSurface Glicoprotein (MSG). MSG is detection target of its presence in the fungus cell wall is very abundant and has a sustainable sequence. The rPCR was successfully optimized with a minimum DNA detection capability of 6.55 cop / μl and did not cross-react with other microorganisms tested in this study. Obtauned 50 clinical sampels consisting of sputum and sputum induction. The result of comparison between microscopic test and rPCR show that rPCR increase positive results up to 20%. The rPCR test can detect P.jirovecii on clinical samples of sputum and induced sputum from HIV patients with pneumonia with CD4 +> 200 or ≤ 200 cells.

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References


Alison M dan Karen AN. (2012). Colonization by Pneumocystis jirovecii and Its Role in Disease. J Clin Microbiol Rev.; 25(2):297-317

Alison M, Kenneth W, Kamyar A, and Laurence H. (2008). Epidemiology and Clinical Significance of Pneumocystis Colonization. J Infect Dis.; 197:10 –7

Anaissie EJ, McGinnis MR, and Pfaller MA.(2009). Clinical Mycology. Churchill Livingstone 1st Edition. Elsevier.

Babic EA, Vilibic CT, Erceg M, Mlinaric ME and Begovac J. 2014. Prevalence of Pneumocystis jirovecii Pneumonia (2010-2013): The First Croatian Report. Acta Microbiol Immunol Hung; 61 (2): 181-8

Calderon EJ, Varela JM, Joly ID, and Cas ED. (2011). Pneumocystis jirovecii Pneumonia. ISBN: 978-1-61209-685-8.

Catharina F, Jacobs JA, Becker P, Templeton KE, Bakkers J, Kuijper EJ. (2006). Interlaboratory Comparison of three Different real time PCR Assay for detection of Pneumocystis jirovecii in Bronchoalveolar Lavage Fluid Samples. J Med Microbiol; 55(pt9): 1229-35

Catherinot E, Lanternier F, Bougnoux ME, Lecuit M, Couderc LJ, and Lortholary O. (2010). Pneumocystis jirovecii Pneumonia. Infect Dis Clin North Am.; 24 (1): 107-38

CDC. Pneumocystis: Pneumocystis jirovecii. DPDx.Unduhhttp://www.cdc.gov/dpdx/pneu mocystis/. 8 agustus 2015

Dohn MN, White ML, Vigdorth EM, Ralph BC, Hertzberg VS, Baughman RP. (2000). Geographic clustering of Pneumocystis carinii pneumonia in patients with HIV infection. Am J Respir Crit Care Med; 162:1617.

Epsy MJ, Uhl JR, SloanLM. 2006. Real-Time PCR in Clinical Microbiology: Application for Routine Laboratory Testing. Clin Microbiol Rev;19(1):165-256

Francisco JM, Marco MC, Conde M, Carmen de la H, Respaldiza N, Antonia G, (2005). Pneumocystis jirovecii in General Population. Emerg Infect Dis.; 11(2): 245-250

Jackop Frans. (2004). Genetic Investigation of Pneumocystis jirovecii: Detection, Cotrimoxazol Resistence and Population Structure. University of Stellenbosch.. Unduh 21 April 2015

Krajicek BJ, Limper AH, and Thomas CF Jr. (2008). Advances in the biology, pathogenesis and identification of Pneumocystis pneumonia. Curr Opin Pulm Med.; 14(3):228-34.

Kutty G, Maldarelli, Achaz G and tin. (2008). The Major Surface Glycoprotein Genes in Pneumocystis jirovecii. J infect Dis; 198(5): 741-749

Munshi A. (2012) DNA Sequencing Methods and Application. ISBN. Unduh http://library.umac.mo/ebooks/b28050393.pdf . 4 Juni 2016

Qiagen. PCR Protocol and Application. https: // www.qiagen.com /resources/ molecularbiology-methods/pcr/. Unduh 3 Juni 2016.

Robert GF, Belaz S, Revest M, Tattevin P, Jouneau S, Decaux O. (2014). Diagnosis of Pneumocystis jirovecii Pneumonia in Immunocompromised Patients by Real-Time PCR: a 4-Year Prospective Study. J Clin Microbiol; 52(9): 3370–3376.

Rodriguez YDA, Wissmann G, Muller AL, Pederiva MA, Brum MC, Brackmann RL. (2011). Pneumocystis jirovecii Pneumonia in Developing Countries. J Parasite.; 18(3): 219228

Rozaliyani A.(2009). Karakteristik klinis, radiologis dan laboratoris pneumonia Pneumocystis pada pasien AIDS dengan gejala pneumonia di beberapa rumah sakit di Jakarta. Jakarta: Medical Research Unit FKUI;.h. 22-51

Stansell JD, Osmond DH, Charlebois E, LaVange L, Wallace JM, Alexander BV. (1997). Predictors of Pneumocystis carinii pneumonia in HIV-infected persons. Pulmonary Complications of HIV Infection Study Group. Am J Respir Crit Care Med.; 155(1): 60-6.

Thermo Scientific. T042 Techmical Bulletin NanoDrop Spectrophotometer: Assesment of Nucleic Acid Purity. Unduh http://www.nanodrop.com/Library/T042NanoDrop-Spectrophotometers-Nucleic-AcidPurity-Ratios.pdf. 2 mei 2016.

Walker J, Corner G, Ho J, Hunt C, and Pickering L. (1989). Giemsa Staining for Cysts and Trophozoites of Pneumocystis carinii. J Clin Pathol; 42(4):432-4




DOI: https://doi.org/10.47007/ijobb.v2i1.22

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