Identifikasi Salmonella sp Pada Jajanan Jus Buah di Kecamatan Gunungpati Semarang dengan PCR

B A S Hermono, S H Bintari, D Mustikaningtyas

Abstract


Berdasarkan hasil observasi dua puskesmas di Kecamatan Gunungpati ditemukan tingginya tingkat kejadian rata-rata penyakit tifus pada tahun 2011-2016. Tujuan penelitian ini mengidentifikasi Salmonella sp pada jajanan jus buah di Kecamatan Gunungpati menggunakan polymerase chain reaction (PCR) dan mengkonfirmasi keberadaan Salmonella sp menggunakan medium triple sugar iron agar (TSIA). Sampel diperoleh dari 17 jajanan jus buah yang dijual di wilayah Kecamatan Gunungpati Semarang. Sampel diisolasi DNAnya menggunakan metode chelex 100-microwave, dilanjutkan PCR dengan primer invA 1 dan invA 2, dan uji konfirmasi menggunakan medium TSIA. Hasil amplifikasi dengan primer invA 1 dan invA 2 menunjukkan tiga sampel jajanan jus buah positif Salmonella sp dengan terbentuknya pita DNA 244 bp. Hasil uji konfirmasi menunjukkan tiga sampel tersebut mengalami perubahan warna medium TSIA dengan kriteria slant berwarna merah, depth kuning, terbentuk gas, dan ada endapan hitam (H2S). Berdasarkan hasil PCR dan konfirmasi kultur pada medium TSIA menunjukkan bahwa 3 dari 17 sampel jus buah positif terkontaminasi Salmonella sp.

Based on the observation from two clinics in Gunungpati Sub District, there was found high rates of thypus disease in the year 2011-2016. The purpose of this research was to identify Salmonella sp inside the fruit juice sold in Gunungpati Sub District by using PCR and to confirm the presence of Salmonella sp using Triple Sugar Iron Agar (TSIA) medium. The samples were collected from 17 fruit juices sold in Gunungpati Sub District, Semarang. The DNA were isolated by chelex 100-microwave methode, continued by testing using PCR with invA 1 and invA 2 primer, and confirmation test using TSIA medium. The result from DNA amplification showed that three samples of fruit juice positively contaminated by Salmonella sp which proved with the formation of DNA strand 244 bp, whereas the result of confirmation test showed that three samples underwent color changing of TSIA medium with the criteria red colored slant,yellow colored depth, the presence of gas, and black precipitation (H2S). Based on PCR test which confirmed by TSIA medium, three out of 17 samples of fruit juices positively contaminated by Salmonella sp


Keywords


Salmonella sp, fruit juices, Gunungpati Sub District

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References


Amalia U. 2013. Optimasi Polymerase Chain Reaction (PCR) untuk Deteksi Salmonella spp. pada Udang Segar. Tesis. Institut Teknologi Bandung

Arum W. 2008. Deteksi bakteri Salmonella pada sampel makanan dan minuman dengan metode Polymerase Chain Reaction. Skripsi. Depok: FMIPA Universitas Indonesia

Bintari SH, Fibriana F, Dewi M, Iswari RS. 2014. PCR approach for rapid detection of Escherichia coli in tempe using a specific primer. J Biological Researches. 19: 54-59

[BPOM] Badan Pengawas Obat dan Makanan Republik Indonesia. 2008. Pengujian mikrobiologi pangan. Journal Info POM 9 (1).

[BSN] Badan Standarisasi Nasional. 2014. SNI. 3719-2014: Minuman sari buah. Jakarta: Badan Standarisasi Nasional

Goldman E, & Lorrence HG. 2015. Practical handbook of microbiology .Third edition. Francis: CRC Press

Lebbofe MJ, & Pierre BE. 2011. A photo graphic atlas for the microbiology laboratory. Morton Publishing Company

Liu CH, & Ou JT. 1996. Rapid identification of Salmonella serovars in feces by specific detection of virulence genes, invA and spvC, by an erichment broth culture-multiplex PCR combination assay. J Clin Microbiol. 34: 2619-1622

Lund BM, Baird-Parker TC, &Gould GW. 2000. The microbiologycal safety and quality of food. Maryland: Aspen Publishers, inc

Malorny B, Hoorfar J, Bunge C, & Helmuth R. 2003. Multicenter validation of the analytical accuracy of Salmonella PCR: Towards an international standard appl environ. Microbiol. 69(1): 290-296

Marwayana ON. 2015. Ekstraksi asam Deoksiribonukleat (DNA) dari sampel jaringan otot. Jurnal oseana. 11(2): 1-9

Prayoga W, & Agustin K.W. 2015. Polymerase Chain Reaction untuk deteksi Salmonella sp. Jurnal Pangan dan Agroindustri. 3(2): 438-488

Rahn K, DeGrandis SA, Clarke RC, McEwen SA, Galan JE, Ginnocch OC, Curtissii R, & Gyles CL.1992. Amplification of invA gene sequence of Salmonella typhimurium by Polymerase Chain Reaction as a specific methode of detection of Salmonella. J Mol Cell Probes. 6: 271-279

Sudian S. 2008. Pengujian mikrobiologi pangan. Jurnal Info POM 9(2): 1-9

Tantri BUN. 2016. Identifikasi bakteri Escherichia coli, Shigella sp, dan Salmonella sp pada air sumur di wilayah pembuangan limbah tahu dan limbah ikan Kota Bandar Lampung. Skripsi. Lampung: Universitas Lampung.

Upadhyay BP, Fuangfa U, Jarinee T, Yuvadee M, Niracha W, & Srisin K. 2010. Detection of Salmonella invA gene in shrim enrichment culture by Polymerase Chain Reaction. J Trop Med Public Health. 41(2): 427-435


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