新型冠状病毒纳米PCR快速检测方法的建立

新型冠状病毒(SARS-CoV-2)感染可导致致命性肺炎,且极具传染性。自2019年年末在我国出现以来,已在全球蔓延。为了建立一种灵敏、快速检测SARS-CoV-2的分子检测方法,本研究使用金纳米颗粒配合普通PCR检测方法,针对SARS-CoV-2核衣壳蛋白建立了SARS-CoV-2 NanoPCR新型分子检测方法。特异性结果显示,该方法对猪流行性腹泻病毒、牛冠状病毒、犬冠状病毒、水貂冠状病毒、猫传染性腹膜炎病毒均无交叉反应,表明该方法的特异性良好。敏感性结果显示,该方法的最低检测量为3.69×10⁴拷贝/μL,高于普通PCR最低检测量10倍,表明该方法的敏感性良好。使用建立的方法对3份临床样品进行检测,该方法与普通PCR方法结果一致,10倍稀释样品后,NanoPCR相比较普通PCR条带仍然具有较高辨识度。综上,本研究建立的灵敏、特异的NanoPCR方法可以对临床样品进行快速诊断和鉴定。

Establishment of a Rapid Detection Method for SARS-CoV-2 by NanoPCR

SARS-CoV-2 infection can cause fatal pneumonia and is extremely contagious.Since it appeared in China at the end of 2019,it has spread globally.We wished to establish a sensitive and rapid molecular-detection method for detection of the nucleic acids(NAs)of SARS-CoV-2.A novel Nano-polymerase chain reaction(PCR)method was established based on a combination of gold nanoparticles and conventional PCR technology with primers designed for detection of the conserved sequences of nucleocapsid proteins.Specificity tested showing that our method had no cross-reactivity with the porcine epidemic diarrhea virus,bovine coronavirus,canine coronavirus,mink coronavirus or feline infectious peritonitis virus:our method had good specificity.The minimum volume of NAs that could be detected using our method was 3.69×10⁴copies/μL,which is 10-times higher than that for conventional PCR:the sensitivity of our method was good.Our method was employed to detect SARS-CoV-2 NAs in three clinical samples,the results of which were consistent with those of conventional PCR.After tenfold dilution of the sample,NanoPCR continued to have a higher degree of recognition than that of conventional PCR bands.In summary,we established a sensitive and specific NanoPCR method to identify SARS-CoV-2 NAs in clinical samples.