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Detection method

A saliva and sample technology, applied in the field of detecting hand, foot and mouth disease in subjects, can solve the problems of time-consuming and long

Pending Publication Date: 2019-11-19
AGENCY FOR SCI TECH & RES +1
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, the above method is rather lengthy and time-consuming (Ooi et al., 2010)
Although rapid diagnostic methods utilizing modern molecular routines such as quantitative real-time PCR (qRT-PCR) have recently been developed to address these issues (Lin et al., 2003), due to multiple genetic differences among enterovirus serotypes, this The sensitivity of this assay needs to be significantly improved (Liang et al., 2012)

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Embodiment 1

[0065] 1. Materials and Methods

[0066] (a). Patient samples and infection

[0067] From August 2012 to February 2016, a total of 35 HFMD suspected throat swabs and saliva clinical samples were obtained from the KK (KK) Women's and Children's Hospital. This collection was approved by the Singapore Health Center Institutional Review Board (CIRB) under CIRB number 2012 / 448 / E. Healthy saliva samples were collected from various children's care centers that participated in the National University of Singapore Ethical Review Board approval number B-14-273 for saliva collection. Confirm the presence of enteroviruses in patient samples using a pan-entero PCR reaction previously established in saliva samples.

[0068] (b). miRNA extraction and reverse transcription

[0069] miRNA was extracted from 50 ul of saliva using Biofluid Extraction Kit (Exiqon, Inc.) and 1 µg of MS2 carrier RNA (Roche, Ltd.), and eluted in 30 µl of water.

[0070] A slightly modified protocol from the manufa...

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Abstract

The present invention relates to a method of detecting Hand-Foot-and-Mouth Disease (HMFD) in subjects. In particular, the method relates to detecting promising biomarkers in the saliva of these subjects. In an aspect of the present invention, there is provided a method for detecting HFMD, the method comprising determining the presence of a microRNA (miRNA) in a saliva sample, wherein the presenceof the miRNA in the saliva sample is indicative of HFMD. Preferably, the miRNA is any one selected from the group consisting of mir-18b, mir-125a, mir-145, mir-155, mir-221, mir-324, mir- 335 and further comprises mir-23a and mir-142.

Description

technical field [0001] The present invention relates to a method of detecting hand, foot and mouth disease (HFMD) in a subject. In particular, the method involves detecting promising biomarkers in the saliva of these subjects. Background technique [0002] HFMD is a widespread viral disease caused by human enterovirus species A (HEV-A) of the genus Enterovirus, afflicting millions of infants and children in the Western Pacific region each year. In recent years, Coxsackievirus A16 (CA16) and enterovirus 71 (EV71) are the main causes of HFMD (Wang and Liu, 2014). However, a surge in the number of cases caused by other HEV-A serotypes such as Coxsackievirus A6 (CA6) has also been reported. HFMD is usually a self-limiting disease characterized by fever and papulovesicular, sometimes maculopapular, palmar, sole, elbow, and trunk rashes, and oral ulcerations (Wang and Liu, 2014). However, EV71-associated HFMD can rapidly develop into severe neurologic complications such as asep...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6851A61K31/7088A61P31/14
CPCA61K31/7088C12Q1/6883C12Q1/701C12Q2600/158C12Q2600/178A61P31/14
Inventor 朱章汉玟·尼奥
Owner AGENCY FOR SCI TECH & RES
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