Development of Dot-blot Hybridization Based on 522 bp Repetitive Sequence (R522) for Detection of Toxoplasma gondii

A. Kusumawati1,4*, H. Widodo2, Tenri Ashari Wanahari3, N. Septiana4 and Sri Hartati5

1Departement of Reproduction, Faculty of Veterinary Medicine, Gadjah Mada University, Jalan sekip unit II, Yogyakarta, 55281 Indonesia. 2Medicinal Plant and Traditional Medicine Research and Developtment Centre, National Institute of Health Research and Development, Ministry of Health Indonesia, Jalan percetakan negara no.29, Jakarta, 10560 Indonesia. 3Faculty of Medicine, Sebelas Maret University, Jalan Ir. Sutami 36 A, Surakarta, 57126 Indonesia. 4Centre of Biotechnology Study, Gadjah Mada University, Jalan teknika Utara, Yogyakarta, 55281 Indonesia. 5Department of Internal Medicine, Faculty of Veterinary Medicine, Gadjah Mada University, Jalan sekip unit II, Yogyakarta, 55281 Indonesia.

ABSTRACT:

Toxoplasmosis, arising from infection by Toxoplasma gondii, is one of the most common parasitic diseases in humans and other warm-blooded animals. In humans, infections are usually asymptomatic but severe disease can occur in immunocompromized individuals and newborns. Due to the importance of the disease and in order to take suitable measures, an early diagnosis of the disease is essential, particularly for pregnant women and in industry of domestic animals. The genome of T. gondii contains repeat sequences B1 and R522 which constitute ideal targets for genome-based detection methods. The 522 base pairs repeat sequences R522 are the most promising due to the high copy number, evaluated to be 200 to 300 units within the genome. We developed a simple dot-blot hybridization based on R522 sequences. The method is simple and does not require sophisticated devices. The test of the method, using cloned R522 as target, showed that the parasite detection method was sensitive and proved to be promising for use in routine health controls as well as for the survey of Toxoplasma infections.

KEYWORDS: DIG-probe; dot blot hybridization; repeat sequences, R522; Toxoplasma gondii

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