Molecular diagnosis of Pneumocystis jirovecii pneumonia by using External PCR in immunocompromised patients .

A. Al-Qaisi, Arkan; A. Al-Khafaji, Alaa; I. Al-Zagroot, Hameed

doi:10.37652/juaps.2014.121391

Document Type : Research Paper

Authors

¹ University of Anbar - College of Medicine

² University of Anbar - College of Pharmacy

10.37652/juaps.2014.121391

Abstract

Pneumocystis jirovecii is an opportunistic eukaryotic pathogen causing life threating pneumonia ( PJP) in immunosuppressed patients.Detection of Pneumocystis jirovecii Pneumonia in immune suppressed patients like hematological cancer with or without chemotherapy, patients on corticosteroid therapy, other malignancy like bronchogenic cancer by using External Polymerase Chain Reaction (External PCR) and to compare the results with other control group (patients who are immunocompetent suffer from pneumonia).A total of 220 specimens were obtained from patients admitted to Al- Ramadi Teaching Hospital, Clinic excitability of respiratory disease in Al- Ramadi and Oncology Teaching Hospital in Baghdad during the period from February to July 2013. This number composed of 71(32%) specimens were from female while the male specimens were 149(68%). A total of 184 (84%) were immunocompromised patients, They were considered immunocompromised patients because they have ( hematological cancer with or without chemotherapy, patients on corticosteroid therapy, other malignancies like bronchogenic cancer). The other 36 (16%) patients with pneumonia were included as immunocompetent patients (No hematological cancer, No chemotherapy, No immunosuppressive drug like corticosteroid therapy and No malignancy like bronchogenic cancer). A total of 220 specimens which were collected, (consisting of 115 blood, 65 sputum, 40 BAL Specimens )the total 220 specimens (115 blood, 65 sputum and 40 BAL Specimens ) were examined PCR technique ( External and Nested PCR ) to identify Pneumocystis Jirovecii DNA based on the amplification of specific primers. The result of agarose gel electrophoresis revealed that small DNA diagnostic bands were detected at 346 bp for External PCR in all positive specimens. Out of 220 specimens 26 (11.8%) were detected by the mtLSU rRNA External PCR. Regarding the use of External PCR technique for sputum specimens 8 (12.3%) were positive, While BAL 8 (20%) and Blood 10 (8.7%) . PCR is more accurate and better indicative for the presence of Pneumocystis jirovecii Pneumonia. In addition this technique can use specimens which were not stored properly and became non suitable for microscopy.

Keywords

Main Subjects

Biology

References

1.Telzak E.; Armstrong D. (1994). Extra pulmonary infection Pneumocystis carinii and other unusual manifestations of Pneumocystis carinii. 361-378.

2.Gajdusek D. (1957). Etiologic agents of interstitial plasma cell pneumonia of premature and infant. Pediatrics. 19:543-565.

3.Neiman P.; Reeves W.; Ray G.; Floumoy N.; Lerner K. G.; Sale G.E.; Thomas E. D. (1976). A prospective analysis of interstitial pneumonia and Pneumocystis carinii pneumonia Pathogenesis, Diagnosis and treatment. 61:326-332.

4.Thomas S.F.; Dutz W.; Khodadad E.J. (1966). Pneumocystis carinii pneumonia (plasma cell pneumonia); roentgen graphic roent·gen·og·ra·phy n. Photography with the use of x-rays. roent gen·o·graph , pathologic and clinical correlation. Am. Roentgen logy roentgenology /roent·gen·ol·o·gy/ (-ol´-ah-je) radiology., Radium Therapy , Nuclear Medicine . J. 98:318-22.

5.Aliouat-Denis C.; Chabé M.; Demanche C.; Aliouat E. M.; Viscogliosi E.; Guillot J.; Delhaes L.; Dei-Cas E. (2008). Pneumocystis species, co-evolution and pathogenic power. Infection Genetics Evolution. 8 (5): 708–726.

6.Zhang J. M.; Zhu J.; Imrich; A.; Cushion M. T.; Kinane B. T.; Koziel H. (2004). Pneumocystis activates human alveolar macrovage NF-KB signaling through mannose receptor . Infect. Immun. 72: 3147-3160.

7.Wakefield A. E.; Pixley F. J.; Banerji S.; Sinclair K.; Miller R. F.; Moxon E. R.; Hopkin J. M. (1990). Detection of Pneumocystis carinii with DNA amplification. Lancet. 336:451–453.

8.Caliendo A. M.; Hewitt P. L.; Allega J. M.; Keen A.; Ruoff K. L.; Ferraro M. J. (1998). Performance of a PCR assay for detection of Pneumocystis carinii from respiratory specimens. Clin. Microbiol. J. 36: 979-982.

9.Thomas C.F.; J.R.; Limper A.H. (2004).Quantifying Pneumocystis jirovecii DNA Med. J. 350:2487–2498.

10.Kanne J. P.; Yandow D. R.; Meyer C. A. ( 2012). Pneumocystis jirovecii Pneumonia: High-Resolution CT Findings in Patients With and Without HIV Infection A.J.R. 198:555-561.

11.Sing A.; Trebesius K.; Roggen kamp A.; Smann H. R.; Tybus K.; Pfaff F.; Bogner J. R.; Emminger C.; Heesemann J.R. (2000). Evaluation of Diagnostic Value and Epidemiological Implications of PCR for Pneumocystis carinii in Different Immunosuppressed and Immunocompetent Patient Groups. Journal Of Clinical Micro. 38(4):1461–1467 .

12.Olsson M.; Elvin K.; Lidman C.; Lofdahls; Linde E. (1996). A rapid and simple nested PCR assay for detection of Pneumocystis carinii in sputum samples. Scand Infect. Dis. J . 28: 597-600.

13.Sheikholeslami F. M.; Sadraei J.; Farina P.; Forozandeh M.; Kochat H. E. (2013). Rate of Pneumocystis pneumonia in Iranian HIV + patients with pulmonary infiltrates. Jundishapur. J .Microbiol. 6 (3):295-300.

14.Leibovitz E.: Pollack H.: Moore T.; Papellas J.; Gallo L.; Krasinski K.; Borkowsky W.(1995). Comparison of PCR and Standard Cytological Staining for Detection of Pneumocystis carinii from Respiratory Specimens from Patients with or at High Risk for Infection by Human Immunodeficiency Virus. Clin .Microbiol. J 33(11):3004-3007.

15.Doskaya M.; Caner A. ; Degirmenci A.; Wengenack N. L.; Yolasigmaz A.; Turgay N.; Toz S. O.; Guruz Y. (2011). Degree and frequency of inhibition in a routine real time PCR detecting Pneumocystis jirovecii for the diagnosis of Pneumocystis pneumonia in Turkey. Journal of Medical Microbiology. 60: 937–944.

16.Govender S.; Plessis S.J.; Ocana G.S.; Chalkley L.J. ( 2008). Prevalence of Pneumocystis jirovecii and Mycoplasma pneumoniae in patients presenting with pneumonia at hospitals in Port Elizabeth. South Afr. J. Epidemiol. Infect. 23(2):21-24.

17.Chen W.; Havell E. A.; Moldawer L. L.; McIntyre K. W.; Chizzonite R. A., Harmsen A. G. (1992). Interleukin 1: an important mediator of host resistance against Pneumocystis carinii. J. Exp. Med. 176:713–718.

18.Beck J. M.; Liggitt H. D.; Brunette E. N.; Fuchs H. J.; Shellito J. E.; Debs R. J. (1991). Reduction in intensity of Pneumocystis carinii pneumonia in mice by aerosol administration of gamma interferon. Infect. Immun. 59:3859–3862.

19.Shellito J.; Suzara V. V.; Blumenfeld W.; Beck J. M.; Steger H. J.; Ermak T. H. (1990). A new model of Pneumocystis carinii infection in mice selectively depleted of helper T lymphocytes. J. Clin. Invest. 85:1686–1693.

20.Yehia M. M.; Al-Habbo D. J. S.; Abdulla Z. A. (2012). Identification and treatment of a patient with Pneumocystis pneumonia (case report). Ann. Coll. Med. Mosul. 38 (1): 68-71.

Journal of University of Anbar for Pure Science

Molecular diagnosis of Pneumocystis jirovecii pneumonia by using External PCR in immunocompromised patients .

References

References

Volume 8, Issue 1
April 2014
Page 1-7

Molecular diagnosis of Pneumocystis jirovecii pneumonia by using External PCR in immunocompromised patients .

References

References

Volume 8, Issue 1April 2014Page 1-7

Volume 8, Issue 1
April 2014
Page 1-7