Volume 8, Issue 2 (4-2020)                   JoMMID 2020, 8(2): 60-64 | Back to browse issues page

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Department of Pathobiology, School of Paraveterinary Sciences, BU-Ali Sina University, Hamadan, Iran.
Abstract:   (520 Views)
Introduction: Cryptococcus neoformans is an encapsulated fungal pathogen that causes life-threatening meningoencephalitis in immunocompromised patients. This yeast secretes several potent immunogenic proteins by secretory vesicular mechanisms, such as HSP70 chaperone. Methods: The PCR-amplified HSP70 gene was cloned into a PET-28a(+) expression vector. The purified recombinant HSP70  (rHSP70) was evaluated by western blotting using an anti-His Tag-HRP antibody and then used for immunization of a rabbit. The serum of the immunized rabbit was tested against the whole lysate of C. neoformans in ELISA. Results: The antibodies in the rabbit’s serum recognized lysate of C. neoformans yeast. The highest antibody levels were achieved after the third booster injection. Conclusion: The rHSP70 showed to be a reliable candidate for the designing and development of an ELISA kit for early detection of cryptococcosis, and to screen a large number of specimens.
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Type of Study: Original article | Subject: Diagnostic/screening methods and protocols
Received: 2020/04/25 | Accepted: 2020/04/13 | Published: 2020/08/16

1. 1. Abassi M, Boulware DR, Rhein J. (2015). Cryptococcal Meningitis: Diagnosis and Management Update, Current Tropical Medicine Reports, 2(2):90-99. [DOI:10.1007/s40475-015-0046-y]
2. Alspaugh JA. (2015). Virulence Mechanisms and Cryptococcusneoformans pathogenesis, Fungal Genetics and Biology, 78:55-58. [DOI:10.1016/j.fgb.2014.09.004]
3. Asea, A, Kaur, P (2018). HSP70 in Human Diseases and Disorders, Springer International Publishing. [DOI:10.1007/978-3-319-89551-2]
4. Bose I, Reese AJ, Ory JJ, Janbon G, Doering TL. (2003). A Yeast under Cover: the Capsule of Cryptococcusneoformans, Eukaryot Cell, 2(4):655-63. [DOI:10.1128/EC.2.4.655-663.2003]
5. Denham ST, Brown JCS. (2018). Mechanisms of Pulmonary Escape and Dissemination by Cryptococcus neoformans, Journal Fungi, 4(1) 1-17. [DOI:10.3390/jof4010025]
6. Eastman AJ, He X, Qiu Y, Davis MJ, Vedula P, Lyons DM, Park YD, Hardison SE, Malachowski AN, Osterholzer JJ, Wormley FL Jr, Williamson PR, Olszewski MA. (2015). Cryptococcal Heat Shock Protein 70 Homolog Ssa1 Contributes to Pulmonary Expansion of Cryptococcusneoformans during the Afferent Phase of the Immune Response by Promoting Macrophage M2 Polarization, Journal of Immunology, 194(12):5999-6010. [DOI:10.4049/jimmunol.1402719]
7. Eileen K. Maziarz, John R. Perfect (2016). Cryptococcosis, Infectious Disease Clinics of North America, 30 (1), 179-206. [DOI:10.1016/j.idc.2015.10.006]
8. Illnait MT, Vilaseca JC, Fernández CM, Martínez GF. (2001). Enzyme-linked Immunosorbent Assay for Detection and Quantification of Cryptococcusneoformans Antigen, Memorias do Instituto Oswaldo Cruz, 96(2):241-245. [DOI:10.1590/S0074-02762001000200018]
9. Kakeya H, Udono H, Maesaki S, Sasaki E, Kawamura S, Hossain MA, Yamamoto Y, Sawai T, Fukuda M, Mitsutake K, Miyazaki Y, Tomono K, Tashiro T, Nakayama E, Kohno S. (1999). Heat shock protein 70 (hsp70) as a major target of the antibody response in patients with pulmonary cryptococcosis, Clinical and Experimental Immunology, 115(3):485-90. [DOI:10.1046/j.1365-2249.1999.00821.x]
10. Khalifeh-Gholi M, Kalali B, Formichella L, Göttner G, Shamsipourd F, Zarnani AH, Hosseini M, Busch DH, Shirazi MH, Markus Gerhard M. (2013). Helicobacter pylori FliD protein is a highly sensitive and specificmarker for serologic diagnosis of H. pylori infection, International Journal of Medical Microbiology, 303(8): 618-623. [DOI:10.1016/j.ijmm.2013.08.005]
11. Perfect JR, Bicanic T. (2015). Cryptococcosis diagnosis and treatment: What do we know now, Fungal Genetics and Biology, 78:49-54. [DOI:10.1016/j.fgb.2014.10.003]
12. Rivet-Dañon D, Guitard J, Grenouillet F, Gay F, Ait-Ammar N, Angoulvant A, Marinach C, Hennequin C.(2015). Rapid diagnosis of cryptococcosis using an antigen detection immunochromatographic test, Journal of Infection, 70(5):499-503. [DOI:10.1016/j.jinf.2014.12.017]
13. Rodrigues ML, Nakayasu ES, Oliveira DL, Nimrichter L, Nosanchuk JD, Almeida IC, Casadevall A. (2008). Extracellular Vesicles Produced by CryptococcusneoformansContain Protein Components Associated with Virulence, Eukaryot Cell, 7(1): 58-67. [DOI:10.1128/EC.00370-07]
14. Rohatgi S and Pirofski LA (2015). Host immunity to Cryptococcusneoformans, Future Microbiology, 10(4), 565-581. [DOI:10.2217/fmb.14.132]
15. Saha DC, Xess I, Zeng WY, Goldman DL. (2008). Antibody responses to Cryptococcusneoformans in Indian patients with cryptococcosis, Medical Mycology, 46, 457-463. [DOI:10.1080/13693780801961345]
16. Silveira CP, Piffer AC, Kmetzsch L, Fonseca FL, Soares DA, Staats CC, Rodrigues ML, Schrank A, Vainstein MH. (2013). The heat shock protein (Hsp) 70 of Cryptococcusneoformans is associated with the fungal cell surface and influences the interaction between yeast and host cells, Fungal Genetics and Biology, 60:53-63. [DOI:10.1016/j.fgb.2013.08.005]
17. Srikanta D, Santiago-Tirado FH, Doering TL. (2014). Cryptococcusneoformans: Historical curiosity to modern pathogen, yeast, 31(2): 47-60. [DOI:10.1002/yea.2997]
18. Upadhya R, Lam WC, Maybruck B, Specht CA, Levitz SM, Lodge JK. (2016). Induction of Protective Immunity to Cryptococcal Infection in Mice by a Heat-Killed, Chitosan-Deficient Strain of Cryptococcusneoformans, MBio, 7(3). [DOI:10.1128/mBio.00547-16]
19. Young M, Macias S, Thomas D, Wormley FL Jr. (2009). A Proteomic-based Approach for the Identification of Immunodominant Cryptococcusneoformans Proteins, Proteomics, 9(9):2578-8. [DOI:10.1002/pmic.200800713]
20. Hashemi SJ, Rezaei S, Ansari S, Daie R, Noorbakhsh F. (2011). Application of Polymerase Chain Reaction (PCR) in the diagnosis of neurocryptoccosis, Tehran University Medical Journal, 69(4): 231-236.