Application Of Cerebrospinal Fluid Next-Generation Sequencing In Diagnosis Of Viral Encephalitis

Objective:To study the application of cerebrospinal fluid(CSF)sequencing technology in the diagnosis of viral encephalitis and evaluate its clinical and laboratory value in accurate diagnosis.Methods:1 ml of CSF was stored in CSF test tube and stored in-20? refrigerator for freezing within 30 min.DNA.were extracted using DNA extraction kit(BGI,Huada Biotechnology Co.,Ltd.,Wuhan,China)for 300 samples of frozen cerebrospinal fluid DNA..the extracted DNA were broken by ultrasound to 200?300 bp size fragments,plus the joint sequence,cyclized to form a single-stranded ring DNA,and then rolled ring amplified by 2-3 orders of magnitude to generate DNA nanospheres.using BGISEQ-50 sequencing platform(huada gene,china)building DNA libraries.the Agilent2100 biological analyzer(AgilentTechnologies,us Santa Clara)was applied to the quality control library insert fragment size,using quantitative PCR(symer fisher tech)quality control DNA library concentration.sequencing was performed using a BGISEQ-50 sequencing platform(tianjin huada institute).Results:Of the 53 patients in the final group,24 were positive by cerebrospinal fluid NGS,including 1 case of pseudorabies virus-causing encephalitis,which was first found and reported in the world,and 11 cases of pseudorabies virus encephalitis were tested in clinical and laboratory tests.The results of cerebrospinal fluid NGS and clinical laboratory tests were statistically analyzed by ?2 test and the McNemar test,it is Statistically significant(P?0.05).Conclusion:1.Cerebrospinal fluid NGS technology has higher sensitivity and has more advantages than traditional laboratory methods in the diagnosis of viral encephalitis.2.cerebrospinal fluid NGS techniques can detect some new or rare pathogenic microorganisms.