应用微滴式数字PCR技术快速诊断新生儿侵袭性真菌病

Clinical value of droplet digital PCR in rapid diagnosis of invasive fungal infection in neonates

摘要:

目的 探讨微滴式数字PCR（ddPCR）技术在快速、精确诊断新生儿侵袭性真菌感染（IFI）中的临床应用价值。方法 选择真菌高度保守序列18S rRNA为目标序列，通过引物设计，建立ddPCR检测真菌体系。在深圳一家医院新生儿重症监护室采集了有IFI高危因素和/或临床症状的83例患者血样（n=83），采用血培养及ddPCR法分别对血样进行真菌检测。结果 ddPCR检测真菌体系的特异性为100%，灵敏度可以达到3.2 copies/μL，重复性良好。临床试验中，22例确诊/临床诊断IFI患儿血样中，ddPCR检测阳性19例。61例拟诊/非IFI患儿血样中，ddPCR法检测结果有2例阳性。结论 初步证明ddPCR技术可用于新生儿IFI的检测，该技术是新生儿IFI筛查甚至诊断很有前途的工具。

Abstract:

Objective To evaluate the clinical value of droplet digital PCR (ddPCR) in rapid and accurate diagnosis of invasive fungal infection (IFI) in neonates. Methods The highly conserved sequence of fungi 18S RNA was selected as the target sequence, and primers were designed to establish a ddPCR fungal detection system. Blood samples were collected from 83 neonates with high-risk factors for IFI and/or related clinical symptoms in the neonatal intensive care unit (NICU) of a hospital in Shenzhen, China. Blood culture and ddPCR were used for fungal detection. Results The ddPCR fungal detection system had a specificity of 100% and a sensitivity of 3.2 copies/μL, and had a good reproducibility. Among the 22 blood samples from neonates with a confirmed or clinical diagnosis of IFI, 19 were detected positive by ddPCR. Among the 61 blood samples from neonates who were suspected of IFI or had no IFI, 2 were detected positive by ddPCR. Conclusions The ddPCR technique can be used for the detection of neonatal IFI and is a promising tool for the screening and even diagnosis of neonatal IFI.