4. Detection of Toxoplasma gondii infection by Polymerase Chain Reaction (PCR) and Histological Examination on Balb/c Mice

Muhammad Hanafiah, Dwinna Aliza, Erdiansyah Rahmi, Wisnu Nurcahyo

Abstract


The purpose of this research was to compare the use of PCR method and histological examination to diagnose toxoplasmosis in tissues of Balb/c mice infected with sporulated oocysts through drinking water. A total of 20 male Balb/c mice aged approximately 2 months were used in this experiment. Each mouse was infected with 1x103Toxoplasma gondii tachyzoites intraperitoneally. Tissue samples (liver, lung, heart, kidney, and brain) were collected from 5 mice on day 1, day 5, day 7, and day 9 after infection. Samples were then examined by PCR and histological methods. The data collected were analyzed descriptively. The results showed that PCR method was more sensitive than histological examination. PCR examination using primer invitrogen gen can amplify DNA T. gondii at 436 bp of the samples from liver, lung, heart and brain on Day 7 and Day 9 after infection. The histological examination showed that the cyst of toxoplasma was found in the brain while mononuclear cells infiltration was found in other internal organs.

Full Text:

PDF

References


Bastien, P., Jumas-Bilak, E., Varlet-Marie, E., and Marty, P. 2007. Three years of multi-laboratory external quality control for the molecular detection of Toxoplasma gondii in amniotic fluid in France. Clin. Microbiol. Infect. 13 (4): 430-433.

Bell, A., and Ranford-Cartwright, L. 2002. Real-time quantitative PCR in parasitology. Trends Parasitol. 18 (8): 338.

Caldearo, A., Piccolo, G., Gorrini, C., Peruzzi, S., Zerbini, L., Bommezzadri, S., Dettori, G., and Chezzi, C. 2006. Comparison between two real-time PCR assays and a nested-PCR for the detection of Toxoplasma gondii. Acta Biomed. 77( 2): 75-80.

Cheng, T.C. 1986. General Parasitology. 2nd Ed. Academic Press College Division, London.

Contini, C., Seraceni, S., Cultrera, R., Incorvaia, C., Sebastiani, A., and Picot, S. 2005. Evaluation of a Real-time PCR-based assay using the light cycler system for detection of Toxoplasma gondii bradizoite genes in blood specimens from patients with toxoplasmic retinochoroiditis. Int. J. Parasitol. 35(3): 275-283.

Dubey, J.P. 2008. The history of Toxoplasma gondii – the first 100 years. J. Eukaryot. Microbiol. 55: 467-475.

Edvinsson, B., Lappalaine, M., and Evengard, B. 2006. Real-time PCR targeting a 529-bp repeat element for diagnosis of toxoplasmosis. Clin. Microbiol. Infect. 12(2): 131-136.

Frenkel, J.K. 1990. Transmission of toxoplasmosis and the role of immunity in limiting transmision and illness, J. Am. Vet. Med. Ass. 196(2): 233-239

Greene, C.E. 2006. Infectious diseases of the dog and cat. 3rd Ed. W.B. Saunders Co., Philadelphia

Grimwood, J. and Smith, J. 1992. Toxoplasma gondii: The role of a 30-kDa surface protein in host cell invasion. Exp. Parasitol. 74: 106-111.

Hanafiah, M., Nurcahyo, W. dan Sumartono. 2003. Studi eksperimental cysts tissue Toxoplasma gondii secara invivo. J. Sain Vet. (2): 27-32.

Lappin, M.R. 1994. Feline Toxoplasmosis. WALTHAM Focus 4(4): 2-8.

Lindstrom, I., Kaddu-Mulindwa, D.H., Kironde, F., and Lindh, J. 2006. Prevalence of latent and reactivated Toxoplasma gondii parasites in HIV-patients from Uganda. Acta Tropica 100: 218-222.

Long, P.L. 1990. Coccidiosis of Man and Domestic Animals. CRC Press. Inc., United States.

Matsuura, T., Tegoshi, T., Furuta-Matsuura, M., and Sugane, K. 1990. Epitope selected monospecific antibodies recombinant antigen from Toxoplasma gondii reacted with dense granules of Tachyoites. J. Histochem. Cytochem. 40: 1725-1730.

McLeod, R., Mack, D., Brown, C. 1991. Toxoplasma gondii-New Advances in Cellular and Molecular Biology. Exp. Parasitol. 63: 272-278.

Montoya, J.G., and Liesenfeld, O. 2004. Toxoplasmosis. Lancet. 363: 1965-1976.

Remington, J. S., and Desmonts, G. 1990. Toxoplasmosis. In: Infectious diseases of the fetus and newborn infant. Remington, J.S. and Klein, J.O. (eds). 3rd Ed. WB. Saunders Company, Philadelphia.

Salant, H.A., Markovics, M.B., Spira, T., Hamburger, J. 2007. The development of a molecular approach for coprodiagnosis of Toxoplasma gondii. Israel Vet. Parasitol. 146: 214–220

Uneke, C.J., Duhlinska, D.D., Nqwu, B.A., and Njoku, M.O. 2007. Seroprevalence of Toxoplasma gondii infection in Kwal, a rural distraction of Plateau-Nigeria. Afr. J. Med. Sci. 36: 109-113.

Xiaohong, H., Xuenan, X., Haruyuki, H., Naoaki, Y., Longshan, X., Naoyoshi, S., and Ikuo, I. 2004. Rapid immunochromatographic test using recombinant SAG2 for detection of antibodies against Toxoplasma gondii in cats. J. Clin. Microbiol. 42: 351-353.

Zhang, H., Thekisoe, O.M., Aboge, G.O, Kyan, H., Yamagishi, J., Inoue, N., Nishikawa, Y., Zakimi, S., and Xuan, X. 2009. Toxoplasma gondii: sensitive and rapid detection of infection by loop-mediated isothermal amplification (LAMP) method. Exp. Parasitol. 122: 47-50.




DOI: https://doi.org/10.21157/ijtvbr.v1i2.6686

Article Metrics

Abstract view : 295 times
PDF - 119 times

Refbacks

  • There are currently no refbacks.


Creative Commons License
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.

 Indexed in: 

Copyright© 2016 | ISSN: 2503-4715 


Published by: 
The Faculty of Veterinary Medicine of Syiah Kuala University
In cooperation with: 
Center for Tropical Veterinary Studies of Syiah Kuala University
and Indonesian Veterinary Medical Association (PDHI)

Online Submissions & Guidelines  | Editorial Policies  | Contact |  Statistics  


Creative Commons License
IJTVBR is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.