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.
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。