Asymptomatic Malaria in Trenggalek District, East Java Province as Revealed by Microscopic Examination (Heny Arwati et al.) 150 ASYMPTOMATIC MALARIA IN TRENGGALEK DISTRICT, EAST JAVA PROVINCE AS REVEALED BY MICROSCOPIC EXAMINATION AND SINGLE STEP PCR Heny Arwati 1 , Kusmartisnawati 1 , Poppy 3 and Yoes Prijatna Dachlan 1, 2 1 Department of Parasitology, Faculty of Medicine, 2 Research Hospital of Tropical and Infectious Diseases, Airlangga University, Surabaya, Indonesia, 3 Pandean Sub-District Community Health Center (Puskesmas), Trenggalek District, East Java Province, Indonesia ABSTRAK Kabupaten Trenggalek merupakan salah satu daerah hipoendemis malaria di Propinsi Jawa Timur. Kasus malaria terbanyak merupakan malaria impor dari daerah endemis di luar pulau Jawa, karena penduduk yang kembali dari bepergian ke daerah tersebut biasanya terinfeksi malaria. Pemeriksaan darah penderita yang sudah dinyatakan sembuh dari malaria diperiksa kembali untuk mengetahui adanya malaria asimtomatis. Diagnosis malaria secara mikroskopis merupakan gold standard yang direkomendasikan oleh WHO. Tetapi karena parasitemia pada malaria asimtomatis sangat rendah dan sulit untuk diperiksa secara mikroskopis, maka diagnosis molekuler dengan single stepPCR dalam penelitian ini digunakan untuk mengkonfirmasi kebenaran diagnosis mikroskopis tersebut. Penelitian ini bertujuan mengetahui adanya malaria asimtomatis di Kecamtan Dongko, Kabupaten Trenggalek secara mikroskopis dan molekular menggunakan metode single step PCR. Pengambilan sampel dilakukan pada bulan Mei dan September 2012. Penentuan responden ditentukan dengan penelusuran data malaria di Puskesmas Pandean Kecamatan Dongko, Kabupaten Trenggalek. Diagnosis secara mikroskopis dilakukan pada sediaan darah tebal dan tipis yang diwarnai dengan Giemsa menggunakan mikroskpe cahaya dengan pembesaran 1000x. Pemeriksaan secara molekular dilakukan dengan metode single step PCR berdasarkan Patsoula et al, (2002). Jumlah sampel darah yang diperoleh dari penderita yang telah sembuh dari malaria adalah 18 sampel. Hasil pemeriksaan mikroskopis adalah 2 sampel terinfeksi P. falciparum adalah, 2 P. vivax, satu campuran dari kedua spesies tersebut dan 13 negatif. Seluruh sampel tersebut kemudian diperiksa dengan single step PCR menghasilkan 2 sampel terinfeksi P. falciparum, 2 P. vivax, 6 campuran kedua spesies tersebut dan 8 sampel sampel negatif. Single step PCR merupakan metode yang lebih akurat dalam mendeteksi dan mengidentifikasi parasit pada malaria asimtomatik dibandingkan dengan cara mikroskopis. Keberadaan malaria asimtomatik merupakan sumber penularan malaria, oleh karena itu deteksi dini dan pengobatan kasus ini perlu dimasukkan dalam program eliminasi malaria khususnya di Kabupaten Trenggalek. (FMI 2013;49:150-154) Kata kunci: malaria asimtomatik, single step PCR, Kabupaten Trenggalek ABSTRACT Trenggalek district is one of hypoendemic areas of malaria in East Java Province. Most of the malaria cases are imported from endemic areas in outside of Java Island, as the destination of the seasonal migratory workers. When they return to Trenggalek district usually they are infected with malaria. Blood examination has been done to the patients who have recovered from malaria infection in order to find out the asymptomatic malaria in this region. The examination of Giemsa-stained blood smear is a gold standard method recommended by WHO. However, due to the very low parasitemia in asymptomatic malaria makes difficulty in microscopic examination, the molecular diagnosis by single step PCR in this research is aimed to confirm the results of microscopic examination of asymptomatic malaria. The objective of this paper was to find out malaria asymptomatic in Dongko sub-district, Trenggalek district by microscopic examination and single step PCR methods. Blood samples were collected during May and September 2012 from the patients who have recovered from malaria infection. The number of sample was according to the list of patients hospitalized in Pandean public health service (Puskesmas Pandean), Dongko sub-district, Pandean district, East Java province. Microscopic diagnosis was done on Giemsa-stained thick and thin smears at 1000x magnification. Molecular diagnosis was done by single step PCR method according to Patsoula et al (2002). During May and September 2012, asymptomatic malaria have been found, where of the 18 samples, 5 samples were positive by microscopic examination two for P. falciparum, two for P. vivax, one for mix of both species, and 13 samples were negative. Single step PCR diagnosed two samples P. falciparum, two samples as P. vivax, 6 samples as mix infection and 6 as negative samples. In conclusion, single step PCR provides a sensitive tool for detection extremely low level of parasitemia in asymptomatic malaria. Mass screening of asymptomatic malaria is needed to be performed simultaneously in collaboration with research institute or research hospital prior to mapping the cases and early medication. The presence of asymptomatic malaria cases is a challenge for the management of malaria elimination program in any endemic area including Trenggalek District. (FMI 2013;49:150-154) Keywords: Asymptomatic malaria, single step PCR, Trenggalek district Correspondence: Heny Arwati, Department of Parasitology, Faculty of Medicine, Airlangga University,