RT-PCR detection of SAP1 gene in Candida albicans isolated from Meningitis cases

Authors

  • Baheeja A. Hmood

DOI:

https://doi.org/10.36320/ajb/v7.i3.8071

Keywords:

Fungal meningitis, Cryptococcus neoformans ,SAP gene

Abstract

Objective :   The main objective of the sampling is to isolate fungi associated with cases of meningitis ,study  of some Virulence factors  of  fungi  and sensitivity to antifungal agents.

Methods: 100 CSF sample were suspected of meningitis culture ,biochemical test and API-System for diagnosis of fungi in sample ,Real-time PCR based on SYBR Green I fluorescent dye for detection of sap gene in Candida albicans.

Results: Depending on agricultures characteristics ,Biochemical tests and  Api  System  the results showed that 32 samples, 32% were given the  positive result for yeast  growth, Candida albicans  found in these samples by 87.5% and Cryptococcus neoformans 12.5% and 25(89.28%) of C.albicans isolates  had Sap gene. C. albicans showed the lowest MICs (1 μg/ml) for amphotericin B. and higher  MICs for  fluconazole (4 - 32 μg/ml) and  Nystatin (8-32 μg/ml) While C.neoformans  showed  low MICs for all antifungal agents  used in this study ranged from (0.06-0.5 μg/ml) for   amphotericin B  and ( 0.25 - 1 μg/ml) for fluconazole and ( 1-4 μg/ml) for nystatin

Conclusion:     Fungal Meningitis which caused by Candida albicans  which have SAP gene that consider as virulence factors  and Resistant to antifungal agents and Cryptococcus neoformans is a debilitating and potentially deadly disease that affects patients with both intact and impaired immune systems. early diagnosis and treatment is essential for optimal outcomes. Patients with Fungal Meningitis have multisystem disturbances and require a well-organized and executed plan of care.

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References

-. National Center for Immunization and Respiratory Diseases(2014) . Fungal meningitis.

- Centers for Disease Control and Prevention (CDC)(2011) " Meningitis: Transmission.

-Ginsberg, L. (2004). "Difficult and recurrent meningitis"). Journal of Neurology, Neurosurgery, and Psychiatry. 75 Suppl 1 (90001): 16–21. DOI: https://doi.org/10.1136/jnnp.2003.034272

-Raman Sharma, R .(2010). "Fungal infections of the nervous system: current perspective and controversies in management". International journal of surgery (London, England) 8 (8): 591–601. DOI: https://doi.org/10.1016/j.ijsu.2010.07.293

- Fernandez M, Moylett EH, Noyola DE, Baker CJ. Candidal meningitis in neonates: a 10-year review. Clin Infect Dis 2000; 31:458. DOI: https://doi.org/10.1086/313973

-Nguyen M.H, Yu ,V.L.(1995). Meningitis caused by Candida species: an emerging problem in neurosurgical patients. Clin .Infect Dis.; 21:323. DOI: https://doi.org/10.1093/clinids/21.2.323

-Njei, B; Kongnyuy, EJ; Kumar, S; Okwen, MP; Sankar, MJ; Mbuagbaw, L (2013). "Optimal timing for antiretroviral therapy initiation in patients with HIV infection and concurrent cryptococcal meningitis.". The Cochrane database of systematic reviews 2: CD009012. DOI: https://doi.org/10.1002/14651858.CD009012.pub2

-Levinson, W. (2010). Review of Medical Microbiology and Immunology (11th ed.). McGraw-Hill.

-Agarwal, N., Kewalramani, N., Kamra, D.N., Agarwal, D.K. and Nath, K. (1991) Hydrolytic enzymes of buffalo rumen: comparison of cell free rumen fluid, bacterial and protozoal fractions. Buffalo Journal 7, 203–207.

-Schaller, M.; Borelli ,C.; Korting, H.C.; Hube, B. (2005) Hydrolytic enzymes as virulence factors of Candida albicans.. Mycoses. 48(6):365-77. DOI: https://doi.org/10.1111/j.1439-0507.2005.01165.x

-Richard, A. Calderone and William, A .Fonzi(2001). Virulence factors of Candida albicans DOI: https://doi.org/10.1016/S0966-842X(01)02094-7

Volume 9, Issue 7, p295-351.

-Julian, R. Naglik1, Stephen J. Challacombe1 and Bernhard Hube2.(2010) Candida albicans Secreted Aspartyl Proteinases in Virulence and Pathogenesis(2010)

-Cauda, R., E. Tacconelli, M. Tumbarello, G. Morace, De, F. Bernardis, A. Torosantucci, and A. Cassone. (1999). Role of protease inhibitors in preventing recurrent oral candidosis in patients with HIV infection: a prospective case-control study. J. Acquir. Immun. Defic. Syndr. 21:20-25. DOI: https://doi.org/10.1097/00126334-199905010-00003

-De Bernardis, F., S. Arancia, L. Morelli, B. Hube, Sanglard, W. Schafer, and A. Cassone. (1999). Evidence that members of the secretory aspartyl proteinase gene family, in particular SAP2, are virulence factors for Candida vaginitis. J. Infect. Dis. 179:201-208. DOI: https://doi.org/10.1086/314546

-Capobianco, J. O., C. G. Lerner, and R. C. Goldman.( 1992). Application of a fluorogenic substrate in the assay of proteolytic activity and in the discovery of a potent inhibitor of Candida albicans aspartic proteinase. Anal. Biochem. 204:96-102. DOI: https://doi.org/10.1016/0003-2697(92)90145-W

-Hube, B. (2000). Extracellular proteinases of human pathogenic fungi, p. 126-137. In J. F. Ernst and A. Schmidt (ed.), Contributions to microbiology. Dimorphism in human pathogenic and apathogenic yeasts. S. Karger AG, Basel, Switzerland. DOI: https://doi.org/10.1159/000060350

-Indrani Bose1, Amy J. Reese1, Jeramia J. Ory1, Guilhem Janbon2, Tamara L. Doering1,(2001). A Yeast under Cover: the Capsule of Cryptococcus neoformans (ASM Press, Washington, D.C.)

- Lee, Mary (2014). Basic Skills in Interpreting Laboratory Data (5 ed.). ASHP. p. 723. Retrieved 16 November.

-Frans ,A. Davied ,D.;Joune, L.(2013).Culture-negative Candida meningitis diagnosed by detection of Candida mannan antigen in CSF. : PubMed,Neurology (Impact Factor: 8.3).; 81(17).

-Rappelli, P., Are, R., Casu, G., Fiori, P. L., Cappuccinelli, P. & Aceti, A. (1998). Development of a nested PCR for detection of Cryptococcus neoformans in cerebrospinal fluid. J Clin Microbiol 36, 3438–3440. DOI: https://doi.org/10.1128/JCM.36.11.3438-3440.1998

- Zaragoza, Oscar Marcio L. Rodrigues, Magdia De Jesus, Susana Frases, Ekaterina Dadachova, and Arturo Casadevall (2009)The capsule of the fungal pathogen Cryptococcus neoformans Adv Appl Microbiol.; 68: 133–216 DOI: https://doi.org/10.1016/S0065-2164(09)01204-0

-Barluzzi R, Brozzetti A, Delfino D, Bistoni F, Blasi E. (1998).Role of the capsule in microglial cell-Cryptococcus neoformans interaction: impairment of antifungal activity but not of secretory functions. Med Mycol.;36(4):189–97. DOI: https://doi.org/10.1080/02681219880000291

-Buchanan, K. L., and J. W. Murphy. (1998). What makes Cryptococcus neoformans a pathogen? Emerg. Infect. Dis. 4:71-83. DOI: https://doi.org/10.3201/eid0401.980109

-Naglik JR, Moyes DL, Wächtler B, Hube B. (2011).Candida albicans interactions with epithelial cells and mucosal immunity. Microbes Infect.;13:963–76. DOI: https://doi.org/10.1016/j.micinf.2011.06.009

-Sanchez-Portocarrero, J., E. Perez-Cecilia, O. Corral, J. Romero-Vivas, and J. J. Picazo. (2000). The central nervous system and infection by Candida species. Diagn. Microbiol. Infect. Dis. 37:169-179. DOI: https://doi.org/10.1016/S0732-8893(00)00140-1

-Mitchell L., Hope Smith S., Braun J.S., Herzog K.H., Weber J.R., Tuomanen E.I. (2004) Dual phases of apoptosis in pneumococcal meningitis. J Infect Dis 190:2039–2046 DOI: https://doi.org/10.1086/425520

-Speed ,B., Dunt, D.(1995).Clinical and host differences between infections with the two varieties of Cryptococcus neoformans. Clin .Infect. Dis.;21(1):28-34. DOI: https://doi.org/10.1093/clinids/21.1.28

-Chuck, S.L, Sande, M.A.(1989) Infections with Cryptococcus neoformans in the acquired immunodeficiency syndrome. N Eng.l J .Med .;321(12):794-9. DOI: https://doi.org/10.1056/NEJM198909213211205

-Sandhu , Stephen ,C. Yixing Li,1 Zundong Yin,1 Zhujun Shao,1 Manshi Li, Xiaofeng Liang, Hardeep S. , . Hadler, Junhong Li, Yinqi Sun, Jing Li, Wenjing Zou, Mei Lin, Shuyan Zuo, Leonard W. Mayer, Ryan T. Novak, Bingqing Zhu, Li Xu, Huiming Luo, (2014). Population-based Surveillance for Bacterial Meningitis in China, Emerg .Infect. Dis. Jan; 20(1): 61–69. DOI: https://doi.org/10.3201/eid2001.120375

-Ahmed, A. Hickey SM, Ehrett S, Trujillo M, Brito F, Goto C, et al.(1996) Cerebrospinal fluid values in the term neonate. Pediatr Infect Dis J.;15:298–303. DOI: https://doi.org/10.1097/00006454-199604000-00004

-Gray, L.D.& Fedorko, D.P.1992, Laboratory Diagnosis of Bacterial Meningitis Clinical Microbiology Reviews Vol.5, No.2 p 130-145. DOI: https://doi.org/10.1128/CMR.5.2.130

-Fitch, M.T., et al.(2008), Emergency department management of meningitis and encephalitis. Infect. Dis. Clin. North. Am,. 22(1): p. 33-52, v-vi. DOI: https://doi.org/10.1016/j.idc.2007.10.001

-Lyons, M.K, Meyer, F.B.(1990). Cerebrospinal fluid physiology and the management of increased intracranial pressure. Mayo Clin Proc.;65:684–707. DOI: https://doi.org/10.1016/S0025-6196(12)65131-3

-. David Gaines, M.D.; Paul B. Eckman, M.D.; Jack S. Remington, M.D. (2009) Low CSF Glucose Level in Fungal Involving the Central Nervous System . Arch Intern Med.;125(2):333-336 DOI: https://doi.org/10.1001/archinte.125.2.333

-Richard, E. Behrman, M.D. (2000). Central Nervous System infection Nelsons textbook of Pediatrics.: 751-752.

- Fishman, R.A.(1992). Cerebrospinal fluid in diseases of the nervous system. 2d ed. Philadelphia: Saunders,.

-Segreti ,J., Harris A.A.(1996). Acute bacterial meningitis. Infect Dis. Clin .North .Am.;10:797–809. DOI: https://doi.org/10.1016/S0891-5520(05)70327-5

-Sakushima, K; Hayashino, Y; Kawaguchi, T; Jackson, JL; Fukuhara, S (2011). "Diagnostic accuracy of cerebrospinal fluid lactate for differentiating bacterial meningitis from aseptic meningitis: a meta-analysis". The Journal of infection 62 (4): 255–62 DOI: https://doi.org/10.1016/j.jinf.2011.02.010

-Zunt ,J.R, Marra, C.M. (1999).Cerebrospinal fluid testing for the diagnosis of central nervous system infection. Neurol Clin.;17:675–89. DOI: https://doi.org/10.1016/S0733-8619(05)70161-4

-Greenlee, J.E.(1990) Approach to diagnosis of meningitis. Cerebrospinal fluid evaluation. Infect Dis Clin North Am.;4:583–98. DOI: https://doi.org/10.1016/S0891-5520(20)30367-6

- Kvaal, C., Lachke, S.A., Srikantha, T., Daniels, K., McCoy, J., and Soll, D.R. (1999) Misexpression of the opaque-phase-specific gene PEP1 (SAP1) in the white phase ofCandida albicans confers increased virulence in a mouse model of cutaneous infection. Infect Immun .67: 6652–6662 DOI: https://doi.org/10.1128/IAI.67.12.6652-6662.1999

-Naglik JR1, Moyes D, Makwana J, Kanzaria P, Tsichlaki E, Weindl G, Tappuni AR, Rodgers CA, Woodman AJ, Challacombe SJ, Schaller M, Hube B. Microbiology.( 2008) Quantitative expression of the Candida albicans secreted aspartyl proteinase gene family in human oral and vaginal candidiasis Nov;154(Pt 11):3266-80. DOI: https://doi.org/10.1099/mic.0.2008/022293-0

-Hube, B., M. Monod, D. A. Schofield, A. J. P. Brown, and N. A. R. Gow. (1994). Expression of seven members of the gene family encoding secretory aspartyl proteinases in Candida albicans. Mol. Microbiol. 14:87-99. DOI: https://doi.org/10.1111/j.1365-2958.1994.tb01269.x

-Leidich, S.D., Ibrahim, A.S. & Fu, Y., et al. (1998) Cloning and disruption of caPLB1, a phospholipase B gene involved in the pathogenicity of Candida albicans. J Biol Chem 273: 26078–26086. DOI: https://doi.org/10.1074/jbc.273.40.26078

-Jitpimolmard S, Sawanyawisuth K, Morakote N, Vejjajiva A, Puntumetakul M, Sanchaisuriya K, Tassaneeyakul W, Korwanich N (2007). Albendazole therapy for eosinophilic meningitis caused by Angiostrongylus cantonensis. Parasitol Res 100: 1293-1296. DOI: https://doi.org/10.1007/s00436-006-0405-7

-Elena ,M. M.; Ulrika, F. T. ;Olivier, B. and Andrej, T. Z.(2012) Activities of Fluconazole, Caspofungin, Anidulafungin, and Amphotericin B on Planktonic and Biofilm Candida Species Determined by Microcalorimetry .

-Arendrup ,M.C; Cuenca-Estrella, M.; Lass-Florl, C., Hope ,W.and Eucast ,A. .( 2012). EUCAST technical note on the EUCAST definitive document EDef 7.2: method for the determination of broth dilution minimum inhibitory concentrations of antifungal agents for yeasts EDef 7.2 (EUCAST-AFST). Clin. Microbiol. Infect. 18:E246–E247. DOI: https://doi.org/10.1111/j.1469-0691.2012.03880.x

-Philpott-Howard, J. N., Wade, J. J., Mufti, G. J., Brammer, K. W. & Ehninger, G. (1993). Randomized comparison of oral fluconazole versus oral polyenes for the prevention of fungal infection in patients at risk of neutropenia. Multicentre Study Group. Journal of Antimicrobial Chemotherapy 31, 973–84. DOI: https://doi.org/10.1093/jac/31.6.973

-John ,E. Bennett, M.D., William E. Dismukes, M.D., Richard J. Duma, M.D., Gerald Medoff, M.D., Merle A. Sande, M.D., Harry Gallis, M.D., John Leonard, M.D., Branch T. Fields, M.D., Major Bradshaw, M.D., Hubert Haywood, M.D., Zell A. McGee, M.D., Thomas R. Cate, M.D., C. Glenn Cobbs, M.D., John F. Warner, M.D., and David W. Alling, M.D., Ph.D.(1979) A Comparison of Amphotericin B Alone and Combined with Flucytosine in the Treatment of Cryptoccal Meningitis.N Engl J Med 1; 301:126-131 DOI: https://doi.org/10.1056/NEJM197907193010303

-Charles M. van der Horst, M.D., Michael S. Saag, M.D., Gretchen A. Cloud, M.S., Richard J. Hamill, M.D., J. Richard Graybill, M.D., Jack D. Sobel, M.D., Philip C. Johnson, M.D., Carmelita U. Tuazon, M.D., Thomas Kerkering, M.D., Bruce L. Moskovitz, M.D., William G. Powderly, M.D., William E. Dismukes, M.D., (1997)Treatment of Cryptococcal Meningitis Associated with the Acquired Immunodeficiency Syndrome. N Engl. J .Med; 337:15-21 DOI: https://doi.org/10.1056/NEJM199707033370103

- Hossain, M. A., G. H. Reyes, L. A. Long, P. K. Mukherjee, and M. A. Ghannoum. (2003). Efficacy of caspofungin combined with amphotericin B against azole-resistant Candida albicans. J. Antimicrob. Chemother. 51:1427-1429. DOI: https://doi.org/10.1093/jac/dkg230

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Published

2015-10-03

How to Cite

A. Hmood, B. (2015). RT-PCR detection of SAP1 gene in Candida albicans isolated from Meningitis cases. Al-Kufa University Journal for Biology, 7(3), 23–35. https://doi.org/10.36320/ajb/v7.i3.8071

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