Potential Point-of-Care Testing for Dengue Virus in the Field

Wang, Wei-Kung; Gubler, Duane J.

doi:10.1128/jcm.00203-18

Cited by 14 publications

(10 citation statements)

References 25 publications

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“…Recently, the portable, simple and compact POCKIT Nucleic Acid Analyzer (POCKIT, GeneReach, Taichung, Taiwan) which can automatically detect and interpret PCR results within one hour became available in mobile PCR laboratory formats [21][22][23]. A lightweight hand-held model, POCKIT Micro Plus Nucleic Acid Analyzer (POCKIT Micro Plus), that works with a built-in rechargeable battery is also available.…”

Section: Rt-pcr [Qrt-pcr])mentioning

confidence: 99%

An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system

et al. 2019

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BackgroundDengue fever, a mosquito-borne disease, is caused by dengue virus (DENV) which includes four major serotypes (DENV-1, -2, -3, and -4). Some serotypes cause more severe diseases than the other; severe dengue is associated with secondary infections by a different serotype. Timely serotyping can provide early warning of dengue epidemics to improve management of patients and outbreaks. A mobile insulated isothermal PCR (iiPCR) system is available to allow molecular detection of pathogens near points of need.Methodology/Principle findingsIn this study, side-by-side comparison with the CDC DENV-1-4 Real Time RT-PCR (qRT-PCR) was performed to evaluate the performance of four singleplex DENV-1–4 serotyping reverse transcription-iiPCR (RT-iiPCR) reagents for DENV subtyping on the mobile PCR system. The four RT-iiPCRs did not react with Zika virus and chikungunya virus; tests with serial dilutions of the four DENV serotypes made in human serum showed they had detection endpoints comparable to those of the reference method, indicating great analytical sensitivity and specificity. Clinical performance of the RT-iiPCR reagents was evaluated by testing 40 serum samples each (around 20 target serotype-positive and 20 DENV-negative); all four reagents had high agreement (97.5–100%) with the reference qRT-PCR. Moreover, testing of mosquitoes separately infected experimentally with each serotype showed that the four reagents detected specifically their target DENV serotypes in mosquito.Conclusions/SignificanceWith analytical and clinical performance comparable to the reference qRT-PCR assay, the four index RT-iiPCR reagents on the field-deployable PCR system can serve as a useful tool for DENV detection near points of needs.

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Section: Rt-pcr [Qrt-pcr])mentioning

confidence: 99%

An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system

et al. 2019

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“…DENV causes different clinical signs in humans, from mild febrile illness (dengue fever, DF) to life-threatening condition such as hemorrhagic fever/dengue shock syndrome (DHF/ DSS) [8,9]. Early, accurate and rapid diagnosis of dengue is critical for confirmation of clinically suspected cases to ensure timely management of severe dengue disease [10]. A number of NS1 rapid diagnostic tests are commercially available to easily detect NS1 antigen, but they are not sensitive or specific enough [11][12][13].…”

Section: Introductionmentioning

confidence: 99%

“…Combined with automated RNA extraction protocols, RT-PCR can potentially facilitate fast and accurate diagnosis of dengue suspected cases with a single sample obtained during the patient's first visit to the clinician. However, the requirement of a pre-PCR nucleic acids extraction step, an expensive and not easy to use thermal cycler, and a highly trained technician make PCR technology not accessible in resource-limited regions where dengue is endemic [10,21]. Molecular diagnosis tools for dengue is still much needed at or near POC/PON, particularly for developing countries with limited public health resources.…”

Section: Introductionmentioning

confidence: 99%

“…Molecular diagnosis tools for dengue is still much needed at or near POC/PON, particularly for developing countries with limited public health resources. For POC/PON dengue diagnosis, several isothermal amplification methods such as nucleic acid sequence-based amplification (NASBA), thermophilic helicase-dependent amplification (tHDA), loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA), and insulated isothermal PCR (iiPCR) have been developed and reported previously as potential field-deployable tools for the detection of different microorganisms [7,10,[22][23][24][25]. Based on these technologies, various all-in-one molecular diagnostics have been made available to help minimizing risks of human errors and allowing easy molecular bio-detection at or near POC/PON in the last few years, but, to our knowledge, no reagents are available for DENV testing on these platforms.…”

Section: Introductionmentioning

confidence: 99%

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A fully automated sample-to-answer PCR system for easy and sensitive detection of dengue virus in human serum and mosquitos

Tsai¹,

Liu²,

Lin³

et al. 2019

PLoS ONE

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Background The insulated isothermal PCR (iiPCR) technology enables consistent PCR amplification and detection in a simple heating device. A pan-dengue virus (DENV) RT-iiPCR, targeting the 5’ untranslated region, was validated previously on the semi-automated POCKIT combo system (involving separate devices for nucleic acid extraction and PCR amplification/detection) to offer performance comparable to a laboratory real-time PCR. Working on the same technologies, a compact automated sample-in-answer-out system (POCKIT Central Nucleic Acid Analyser) has been available commercially for iiPCR, minimizing human error risks and allowing easy molecular bio-detection near points of need. Here, we evaluated the analytical and clinical performance of the pan-DENV RT-iiPCR on the fully automated system by comparison to those on the semi-automated system. Methodology/Principal findings Testing sera containing serial diluted DENV-1, -2, -3, or -4 cell culture stock, the pan-DENV RT-iiPCR system had similar 100% detection endpoints on the two systems; i.e. at 1, 10, 1 and 10 PFU/ml, respectively, on the fully automated system, and at 10, 1, 10 and 10 PFU/ml, respectively, on the semi-automated system. Furthermore, both fully automated and semi-automated PCR system can detect all four DENV serotypes in mosquitos. Clinical performance of the reagent on the two systems was evaluated by testing 60 human serum samples. Both systems detected the same 40 samples (ten DENV-1, -2, -3, and -4 positive each) and did not detect the other 20; 100% agreement (κ = 1) was found between the two systems. Conclusions/Significance With performance comparable to a previously validated system, the fully-automated PCR system allows applications of the pan-DENV reagent as a useful tool near points of need to facilitate easy, fast and effective detection of dengue virus and help mitigate versatile public health challenges in the control and management of dengue disease.

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“…As of recently, there are no specific vaccines to treat dengue except the CYD-TDV (Dengvaxia, Sanofi Pasteur), the first licensed dengue vaccine that is approved by the U.S. Food and Drug Administration (FDA). However this vaccine is compromised by several limitations such as being restricted to patients who have experienced previous DENV infection [3][4].…”

Section: Introductionmentioning

confidence: 99%

Recent Trends in Dengue Detection Methods Using Biosensors

Zainuddin

Nordin

Rahim

2018

IIUMEJ

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Dengue illness is an infectious tropical disease, transmitted by Aedes mosquitos, that poses a serious health threat to the tropical world. This disease causes widespread infection worldwide, with about 50 million cases of dengue occurring per annum out of which 500,000 recorded cases of dengue hemorrhagic fever and 22,000 deaths. Currently, there are no effective vaccines available to prevent the spread of the infection. Accurate and rapid laboratory diagnostic tests are required for early detection to reduce patient mortality rate. In this paper, common laboratory diagnosis methods for detecting dengue virus infection are discussed. Currently, virus isolation, RT-PCR and serology methods provide the most direct and accurate response for detection of dengue. However, these methods require tedious steps, expensive requirements and expert staffs. Recent research have proposed the usage of biosensors as an alternative new technology for detection of dengue. In this work, various types of biosensors such as electrochemical, piezoelectric, and optical biosensor have been described and compared to evaluate their effectiveness in dengue detection. It is observed that the optical biosensor offers the best detection due to its high sensitivity as compared to others, although it is popularly known as an expensive method. Alternatively, the use of electrochemical and piezoelectric biosensors (QCM) is highly recommended for detection of dengue due to their ease-of-use, low cost, low reagent consumption, disposability, and minimal sample preparation. These approaches have the potential to improve the rate of survival, particularly in resource-limited countries. ABSTRAK: Virus denggi adalah penyakit berjangkit tropika bawaan nyamuk Aedes yang menimbulkan ancaman serius kepada kesihatan global. Penyakit ini menyebabkan jangkitan yang meluas di seluruh dunia, dengan kira-kira 50 juta kes denggi yang berlaku setiap tahun di mana 500,000 kes demam denggi berdarah dan 22,000 kematian direkodkan. Buat masa ini, tiada vaksin yang berkesan untuk mencegah penyebaran jangkitan ini. Ujian diagnostik makmal yang cepat dan tepat diperlukan untuk pengesanan denggi awal untuk mengurangkan kadar kematian pesakit. Dalam artikel ini, kaedah diagnosis makmal yang biasa dilakukan untuk mengesahkan jangkitan virus denggi akan dibincangkan. Pada masa kini, kaedah pengasingan virus, RT-PCR dan serologi adalah tindak balas yang paling cepat dan tepat untuk mengesan denggi. Walau bagaimanapun, kaedah-kaedah ini memerlukan langkah-langkah yang melecehkan, kos penyelenggaraan yang tinggi dan kakitangan yang terlatih. Penyelidikan terkini telah mencadangkan penggunaan biosensor sebagai teknologi baru alternatif untuk mengesan denggi. Dalam artikel ini juga pelbagai jenis biosensor seperti biosensor elektrokimia, piezoelektrik, dan biosensor optik telah dijelaskan dan dibandingkan untuk menilai keberkesanannya dalam pengesanan denggi. Difahamkan bahawa biosensor optik menawarkan pengesanan terbaik kerana kepekaannya yang tinggi berbanding dengan yang lain, walaupun ia dikenali sebagai kaedah berkos tinggi. Sebaliknya, biosensor elektrokimia dan piezoelektrik (QCM) sangat disyorkan untuk mengesan denggi kerana ia mudah digunakan, berkos rendah, penggunaan bahan uji yang terhad, boleh dipakai buang, dan mempunyai penyediaan sampel yang minima. Pendekatan-pendekatan ini berpotensi untuk meningkatkan kadar kemandirian di kawasan-kawasan sumber terhad.

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Potential Point-of-Care Testing for Dengue Virus in the Field

Cited by 14 publications

References 25 publications

An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system

An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system

A fully automated sample-to-answer PCR system for easy and sensitive detection of dengue virus in human serum and mosquitos

Recent Trends in Dengue Detection Methods Using Biosensors

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