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. 2019 Jan;21(1):45-51.
doi: 10.7499/j.issn.1008-8830.2019.01.009.

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

[Article in Chinese]
Hui-Tao Li  1 , Bing-Chun Lin, Zhi-Feng Huang, Chuan-Zhong Yang, Wei-Min Huang
Affiliations

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

[Article in Chinese]
Hui-Tao Li et al. Zhongguo Dang Dai Er Ke Za Zhi. 2019 Jan.

Abstract
in English, Chinese

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。

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Figures

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微滴式数字PCR法梯度温度扩增白假丝酵母菌基因组DNA的一维散点图（左）及最佳温度（59.2℃）下的峰图（右）
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微滴式数字PCR法梯度稀释扩增白假丝酵母菌基因组DNA的一维散点图
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微滴式数字PCR法梯度扩增3株真菌基因组DNA及10株细菌基因组DNA的一维散点图
See this image and copyright information in PMC

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