Detection of Enterovirus Genotypes by CRISPR Technology

Children's Hospital of Fudan University

Overview

CRISPR-enterovirus detection system was constructed in this study for detection variety genotypes of enterovirus rapidly in children suspected or diagnosed as enterovirus infection.

To construct CRISPR-enterovirus detection system, we screened the targets, primers and CrRNA and evaluated the capability of this detecting system. Samples of feces, blood and cerebrospinal fluid (the remainder of routine testing samples) from 500 children with suspected or confirmed enterovirus infection were examined using the CRISPR technique. To evaluate the CRISPR-enterovirus detection system, real-time PCR and traditional PCR as comparison method was implemented. The method of real-time PCR and traditional PCR were used to detect enterovirus and analyze the genotypes of enterovirus, respectively. According to the results of CRISPR-enterovirus detection system and the comparison method, the positive coincidence rate, negative coincidence rate and consistency of the two methods were calculated. Furthermore, the sensitivity and specificity of the CRISPR-enterovirus detection system for detection of enterovirus were calculated.

Study Type

OBSERVATIONAL

Conditions

Enterovirus Infections

Interventions

Non-invasive detection method: CRISPR techonologyOTHER

This study was a laboratory diagnostic study without any intervention.

Eligibility

Sex: ALLMin age: 1 DayMax age: 18 Years

Medical Language ↔ Plain English

Inclusion Criteria: * Patients with clinically suspected or confirmed enterovirus infection Exclusion Criteria: * Patients with clinical diagnosis of non-enterovirus infection

Outcomes

Primary Outcomes

Successful construction of CRISPR-enterovirus detection system

It is a binary variable ("yes/no").If the system meet all the following criteria, it would be setted into "yes" .According to the results of CRISPR-enterovirus detection system and the comparison method： 1. the positive coincidence rate and negative coincidence rate are both over 95.0%; 2. the value of Kappa using to express the consistency of those two methods is over 0.75; 3. the sensitivity and specificity of the CRISPR-enterovirus detection system for detection of enterovirus are over 90.0% and 95.0%; 4. the coefficient of variation between batches is less 15.0%, and each batch detection takes less than 1 hour.

Time frame: At enrollment

Data from ClinicalTrials.gov

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