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Non-Commercial Use Only Infectious Disease Reports 2020; volume 12(s1):8731 Performance comparison of higher (100 %) when using real-time PCR. two malaria rapid diagnostic Both RDTs showed a 100% agreement. RT- Correspondence: Puspa Wardhani, PCR detected higher mix infection when Department of Clinical Pathology, Faculty of test with real time polymerase compared to microscopy and RDTs. Medicine, Universitas Airlangga/Dr. Soetomo. chain reaction and gold Conclusion: RightSign and ScreenPlus General Academic Hospital, Jl. Prof. Dr. standard of microscopy RDT have excellent performance when Moestopo No. 47, 60132, Surabaya, using microscopy detection as a gold stan- Indonesia. detection method dard. Real-time PCR method can be consid- Tel: +628123176937 E-mail: [email protected], ered as a confirmation tool for malaria diag- 1,5 Puspa Wardhani, Trieva Verawaty nosis. Key words: Malaria, rapid diagnostic test, 2,5 Butarbutar, Christophorus Oetama real-time polymerase chain reaction, Adiatmaja,2,5 Amarensi Milka microscopy detection. Betaubun,3,5 Nur Hamidah,4 Aryati1,5 Contributions: Conceptualization: PW. Data 1Clinical Pathology Department; Introduction curation: PW TVB COA AMB. Formal analy- 2 Indonesia is a malaria endemic area. Clinical Pathology Specialist Study sis: PW AA NH. Funding: KEMENRIS- Programme, Clinical Pathology The province of Papua has Indonesia’s TEKDIKTI RI. Investigation: PW PW TVB Department; 3Clinical Pathology highest malaria burden. All Plasmodium COA AMB, NH, AA. Methodology: PW AA. Subspecialist Study Programme, species are present in Papua, including the Project administration: PW TVB AMB COA Plasmodium knowlesi (P. knowlesi) Clinical Pathology Department, Faculty Resources: PW AA NH. Supervision: PW AA originally discovered on Kalimantan Island. NH. Validation: PW AA. Writing–original of Medicine; 4Science and Technology The most common types of Plasmodium draft, review & editing: PW. Faculty, C Campus, Universitas 5 infection in Papua are Plasmodium Airlangga; Dr. Soetomo General falciparum (P. falciparum) and Plasmodium Conflicts of interest: The authors declare no Academic Hospital, Surabaya, Indonesia vivax (P. vivax). Plasmodium ovale (P. potential conflict of interest. ovale) and Plasmodium malariae (P. only malariae) also can be found in Papua. The Funding: The work was supported by Simlitabmas Programme by Kementretian highest cause of morbidity and mortality in 1,2 Riset Teknologi dan Pendidikan Tinggi Abstract Papua is P. falciparum. Republik Indonesia (KEMENRISTEKDIKTI Several methods for diagnosing malariauseRI), contract number: 544/UN3.14/LT/2019. Background: The diagnostic test for have been established since WHO has malaria is mostly based on Rapid confirmed how important new tests are to Acknowledgements: The authors acknowl- Diagnostic Test (RDT) and detection by diagnose Plasmodium spp rapidly, reliably, edge the funding research support of microscopy. Polymerase Chain Reaction accurately and cheaply, to address Simlitabmas programme and Merauke (PCR) is also a sensitive detection method numerous shortcomings in microscopic Hospital, Papua for sampling place support, that can be considered as a diagnostic tool. examination as the WHO’s gold standard Parasitology department Faculty of Medicine, The outcome of malaria microscopy detec- for malaria research.3,4 Brawijaya University for consultation in labo- tion depends on the examiner’s ability and Microscopic examination has ratory method. experience. Some RDT has been distributed advantages, namely allowing to identify Conference presentation: The paper had been in Indonesia, which needs to be evaluated species definitively, determine the condition presented at INSBIOMM International confer- for their results. of parasitemia, monitor malaria treatment ence on latest perspectives on Infectious Objective: This study aimed to compare response, easy execution and inexpensive3,5. the performance of RightSign RDT and Diseases, including Biotreats and Military However, the weakness of microscopy Medicine, in August 27-28th, 2019, Surabaya, ScreenPlus RDT for detection of examination is the difficulties in detecting INDONESIA. Plasmodium in human blood. We used spe- extrimely low parasitemia and mix cific real-time polymerase chain reaction infections, and it is time consuming. Received for publication: 17 February 2020. abTESTMMalaria qPCRII) and goldNon-commercial stan- Although microscopy examination is gold Accepted for publication: 1 July 2020. dard of microscopy detection method to standard in malaria diagnosis, it is better to measure diagnostic efficiency. be accompanied by RDTs or other This work is licensed under a Creative Methods: Blood specimens were evalu- methods.3,5 Commons Attribution-NonCommercial 4.0 International License (CC BY-NC 4.0). ated using RightSign RDT, ScreenPlus Other testing methods have been RDT, Microscopy detection, and RT-PCR developed to diagnose malaria are the based ©Copyright: the Author(s), 2020 as the protocol described. The differences on proteins produced by Plasmodium, such Licensee PAGEPress, Italy on specificity (Sp), sensitivity (Sn), positive as Histidine-Rich Protein 2 (HRP-2) or Infectious Disease Reports 2020; 12(s1):8731 predictive value (PPV), and negative pre- enzymes such as Pan-Plasmodium Lactate doi:10.4081/idr.2020.8731 dictive value (NPV) were analyzed using Dehydrogenase (pLDH) and Pan-Specific McNemar and Kruskal Wallis analysis. Aldolase. This method has been widely Results: A total of 105 subjects were used and is considered an alternative to recruited. Based on microscopy test, malaria testing. This examination is also examination tools generally use serological RightSign RDT had sensitivity, Specificity, very prominent, especially in areas where techniques to detect antigens, such as Rapid PPV, NPV, 100%, 98%, 98.2%, 100%, microscopic examination is not available, or Diagnostic Tests (RDTs) or respectively. ScreenPlus showed 100% it is available but there is no laboratory staff Immunochromatography Test (ICT) and sensitivity, 98% specificity, 98.2% PPV, experienced in the examination of blood or Enzyme Linked Immunosorbent Assay 100% NPV. The sensitivity of both RDTs some other disadvantage of microscopy (ELISA)4,6,7. became lower (75%) and the specificity examination. Commercially enzyme-based The RDTs or ICT detect malaria [page 56] [Infectious Disease Reports 2020; 12(s1):8731] Article antigens when the blood of the patient Sample collection results was carried out by two blind and passes through a membrane containing This study was a cross-sectional analyt- independent observers. Based on the Plasmodium-specific antibodies. The ic study conducted at Merauke Hospital, McNemar test between the two observers’ benefits of RDT are reliability in testing, Papua and Clinical Pathology Laboratory, results, the results were not significantly quick processing times and tests, and cheap Faculty of Medicine, Universitas Airlangga different with P<0.05. review fees, which is why it is very useful /Dr. Soetomo General Academic Hospital in endemic areas. The most commonly used Surabaya during November 2018-June Real Time PCR RDTs are methods of detection based on 2019. A total of 105 whole blood samples DNA examination of Plasmodium HRP2 and pLDH. The advantages of RDTs were collected in tubes containing Ethylene malaria using the Rotor-Gene® Q PCR are their parasitic detection limit of 50-100 Diamine Tetraacetic Acid (EDTA). from Qiagen, Tokyo, Japan with the parasites/μL. The disadvantage of RDT are abTESTMMalaria qPCR III reagent kit. reduced effectiveness and efficiency in Species identification and determi- AbTESTMMalaria qPCR III (AITbiotech examining large samples, because time nation of parasite density Pte Ltd, Singapore) can identify four consuming and a high level of Identification of Plasmodium species species of Plasmodium spp. (P. falciparum, concentration and accuracy are required.3, 8 and calculation of the parasitemia index P. vivax, P. ovale, P. malariae, P.knowlesi). Most RDTs examine two proteins at (PI) were performed microscopically on There are two main steps: DNA extraction once, such as a combination of a specific Giemsa-stained thick and thin blood from blood samples and amplification of protein P. falciparum (HRP-2) with pLDH, smears. PI was calculated using WHO DNA extracts using primers pairs and a protein that is shared by all species of guidelines Simultaneously the samples probes that hydrolyze double-dye (double- Plasmodium (non-specific) pLDH) or HRP- were using RightSign RDT and ScreenPlus dye hydrolysis probes) which are very 2 combination comnined with a species RDT.12 specific. Double-dye hydrolysis probes are specific protein species of Plasmodium fluorescent substances that are able to emit vivax (Pv-pLDH). Three proteins also can RightSign RDT and ScreenPlus light, then the light will be captured by be detected at once, such as HRP-2, non- RDT optical detectors on the Rotor-Gene® Q device, which results in an increase in specific pLDH and Pv-pLDH that can The positivity and antigen detection of only signal in the wave graph. This probe distinguish between falciparum malaria and the Plasmodium species in RightSign RDT 9 attaches to the primers of each Plasmodium non-falciparum malaria or mixed malaria. were obtained through lines or bands that species, which are compatible with the RightSign and ScreenPlus are available arise in the line test. RightSign uses the Rotor-Gene® Q optical detector channel, comercially in Indonesia. RightSign detects immunochromatography
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