DNA Quality Testing of Isolated Stored Giemsa Blood Smear Samples using Geneius™ Micro gDNA Extraction Kit as An Alternative Sample for Malaria Molecular Examination
Abstract
The gold standard for malaria examination is microscopic examination. However, microscopic examination has many shortcomings, namely the difficulty of distinguishing between Plasmodium sp. species when mixed infections occur. Other malaria examinations for screening can be done with RDT. However, it also has shortcomings, namely the minimum number of parasites that can be detected is 100 parasites/ul. Molecular examination is the most sensitive and accurate examination in the diagnosis of malaria. The initial stage of molecular examination is the DNA isolation stage. This DNA isolation is very important in determining the success of molecular examination. Isolation of Plasmodium sp. DNA using blood smear samples is still rarely done and there is no standard procedure for isolating DNA from blood smears. The purpose of this study was to determine whether stored Giemsa blood smears can be used as samples in DNA isolation in malaria cases. The method used in isolation is using the Geneius™ Micro gDNA Kit. The results of the study were that stored Giemsa blood smear samples of Plasmodium vivax for 3 months could be isolated using the Geneius™ Micro gDNA Kit and could be used as DNA templates for amplification with target gene primers.
References
Adel, S., Saeed, M., & Ali, P. (2014). Nested Polymerase Chain Reaction (PCR) on Fixed Stained Slides in Comparison to Whole Blood as a Source of DNA in Southeast of Iran. J Trop Dis, 2(3).
Aschar, M., Sanchez, M. C. A., Costa-Nascimento M, dJ, & Farinas, M. de LR. (2022). Ultrasensitive molecular tests for Plasmodium detection: applicability in control and elimination programs and reference laboratories. Rev Panam Salud Publica, 46.
Ashley, E. A., Pyae Phyo, A., & Woodrow, C. J. (2018). Malaria. Lancet, 21, 1608–1621.
Bastiaens, G., Bousema, T., & Leslie, T. (2014). Scale-up of malaria rapid diagnostic tests and artemisinin- based combination therapy: challenges and perspectives in sub-Saharan Africa. PLoS Med, 11(1).
Calderaro, A., Piccolo, G., & Chezzi, C. (2024). The Laboratory Diagnosis of Malaria: A Focus on the Diagnostic Assays in Non-Endemic Areas. Int. J. Mol. Sci, 25(695).
CDC. (2013). Malaria. Http://Www.Cdc.Gov/Dpdx/Malaria/.
Donastin, A., Koentjoro, M. P., Hidayat, M. T., & Prasetyo, E. N. (2022). Perbandingan Tiga Metode Isolasi DNA Aspergilus niger. Jurnal Metamorfosa, 9(1), 67–78.
Hutami, R., Bisyri, H., Sukarno, S., Nuraini, H., & Ranasasmita, R. (2018). Ekstraksi DNA dari Daging Segar untuk Analisis dengan Metode Loop-Mediated Isothermal Amplification (LAMP). Jurnal Agroindustri Halal, 4(2), 209–216.
Inderiati, D., Handayani, S., Syaeptiani, D., & Aryadnyani, N. P. (2022). Identifikasi Plasmodium vivax Menggunakan Metode Nested PCR Di Wilayah Endemis Malaria Provinsi Nusa Tenggara Timur. Journal of Indonesian Medical Laboratory and Science (JoIMedLabS), 3(1), 38–50.
Mann, R., Sharma, S., Mishra, N., Valecha, N., & Anvikar, A. R. (2015). Comparative assessment of genomic DNA ectraction processes for Plasmodium: identifying the appropriate method. Journal Vector Borne, 52, 273–280.
Mustafa, H., Rachmawati, I., & Udin, Y. (2016). Genomic DNA concentration and purity measurement of Anopheles barbirostris. Journal of Disease Vector, 10(1), 7–10.
Panda, B. B., Meher, A. S., & Hazra, R. K. (2019). Comparison between different methods of DNA isolation from dried blood spots for determination of malaria to determine specificity and cost effectiveness. Journal of Parasitic Diseases, 43(3), 337–342.
Perwitasari, D. A., Faridah, I. N., Ratnasari, Y. A., Agustina, K., Utami, I. N., & Maliza, R. (2020). Comparison of DNA Isolation Methods using FTA Card: Wizard® Kit Genomic DNA Purification, PureLink® Genomic DNA, and Chelex-100. Jurnal Ilmu Kefarmasian Indonesia, 18(2), 241–245.
Puasa, R., & Kader, A. (2018). Identifikasi Plasmodium Malaria Didesa Beringin Jaya Kecamatan Oba Tengah Kota Tidore Kepulauan. Jurnal Riset Kesehatan, 7(1), 21–24.
Putri, N. P. P. E., & Junitha, I. K. (2015). Kualitas dan Kuantitas DNA Darah Kering Pada Besi Dan Kayu Yang Disimpan Dalam Kurun Waktu Berbeda. Jurnal Biologi , 19(1), 21–24.
Restu, M., Mukrimin, M., & Gusmiaty, G. (2012). Optimalisasi teknik ekstraksi dan isolasi DNA tanaman suren (Toona Sureni Merr.) untuk analisis keragaman genetik berdasarkan Random Amplified Polymorphic DNA (RAPD). Jurnal Natur Indonesia, 14(1), 138–142.
Sambe, B. S., Diagne, A., Diatta, H. A. W., Gaba, F. M., Sarr, I., Diatta, A. S., Ousmane, S., Diaw, M., Sane, R., Diouf, B., Vigan-Womas, I., Mbengue, B., & Niang, M. (2023). Molecular detection and quantification of Plasmodium vivax DNA in blood pellet and plasma samples from patients in Senegal. Frontiers in Parasitology, 2:1149738.
Seesui, K., Imtawil, K., Chanetmahun, P., Laummaunwai, P., & Boonmars, T. (2018). An alternative method for extracting Plasmodium DNA from EDTA whole blood for malaria diagnosis. Korean J Parasitol, 55(1), 25–32.
Sirait, P. S., & Sulistiawati, S. (2023). Pengaruh Perbedaan Metode Isolasi Terhadap Kualitas DNA Ikan Gabus (Ophiocephalus sp.). Journal of Global Sustainable Agriculture, 4(1), 96–100.
Syafaruddin, S., & Santoso, T. J. (2015). Optimasi Teknik Isolasi dan Purifikasi DNA yang Efisien dan Efektif pada Kemiri Sunan (Reutalis trisperma (Blanco) Airy Shaw). Jurnal Penelitian Tanaman Industri, 17(1).
Syaifudin, M., Wiluyaningtias, D. H., & Yuliwati. (2015). Evaluation of five methods used to extract deoxyribonucleic acid (DNA) from human malaria parasitized blood spotted on the filter paper. Asian Journal of Applied Sciences, 03(03), 429–436.
Wang, B., Han, S., Cho, C., Han, J., Cheng, Y., Lee, S., Galappaththy, G. N. L., Thimasarn, K., Soe, M. T., Oo, H. W., Kyaw, M. P., & Han, E. (2014). Comparison of microscopy, nested-PCR, and Real-Time-PCR assays using high-throughput screening of pooled samples for diagnosis of malaria in asymptomatic carriers from areas of endemicity in Myanmar. Journal of Clinical Microbiology, 52(6), 1838–1845.
Wasdili, F. A. Q., Putri, A. S., & Romlah, S. (2022). Optimasi Isolasi DNA Genom Candida albicans dengan Metode Litium Karbonat. Anakes : Jurnal Ilmiah Analis Kesehatan , 8(2), 160–168.
WHO. (2014). World Malaria Report 2014. WHO Press.
WHO. (2017). WHO-FIND Malaria RDT Evaluation Programme: Product Testing Round 7. WHO Press.
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