结直肠癌组织miR-137-3p、miR-410-3p表达水平与上皮间充质转化和预后的关系分析

摘要 目的：探讨结直肠癌（CRC）组织微小RNA-137-3p（miR-137-3p）、微小RNA-410-3p（miR-410-3p）的表达与上皮间充质转化（EMT）和预后的关系。方法：选取2017年2月至2019年2月本院收治的115例接受CRC根治术治疗的患者，采用实时荧光定量聚合酶链式反应（qRT-PCR）检测癌组织及癌旁组织中miR-137-3p、miR-410-3p表达、EMT标志物E-钙粘蛋白（E-cadherin）、波形蛋白（Vimentin）的mRNA表达并进行相关性分析，分析癌组织中miR-137-3p、miR-410-3p表达与CRC临床病理特征的关系。术后随访3年，采用Kaplan-Meier法分析miR-137-3p、miR-410-3p高表达和低表达患者的预后情况。结果：癌组织中miR-137-3p表达及E-cadherin mRNA表达水平低于癌旁组织，miR-410-3p表达及Vimentin mRNA表达水平高于癌旁组织（P＜0.05）。癌组织中miR-137-3p表达与E-cadherin mRNA表达水平、miR-410-3p表达与Vimentin mRNA表达水平分别呈正相关（P＜0.05），癌组织中miR-137-3p表达与Vimentin mRNA表达水平、miR-410-3p表达与E-cadherin mRNA表达水平分别呈负相关（P＜0.05）。miR-137-3p、miR-410-3p表达与TNM分期、肿瘤分化程度、淋巴结转移有关（P＜0.05）。Kaplan-Meier生存曲线分析显示，miR-137-3p低表达、miR-410-3p高表达患者3年累积生存率下降（P＜0.05）。结论：CRC组织中miR-137-3p表达下调、miR-410-3p表达上调，miR-137-3p、miR-410-3p表达水平与EMT密切相关，且miR-137-3p高表达、miR-410-3p低表达患者的预后更好。     

MicroRNA-520e通过靶向抑制AEG-1发挥抗结直肠癌特性

摘要 目的：探究MicroRNA-520e（miR-520e）在结直肠癌中的表达模式及其对细胞功能的影响。方法：采用qRT-PCR方法检测47例结直肠癌患者的癌组织和癌旁组织中miR-520e和星形胶质细胞上调基因-1（AEG-1）的mRNA表达水平。将SW480细胞分为对照组、miR-520e-mimic组、NC-mimic组、miR-520e-inhibitor组、NC-inhibitor组、miR-520e-mimic+AEG-1-pcDNA3.1组和miR-520e-mimic+NC-pcDNA3.1组。通过MTT法检测SW480细胞的增殖,通过Annexin V-FITC/PI双染色试剂盒检测细胞凋亡,通过Transwell检测细胞迁移和侵袭,通过双荧光素酶报告基因实验验证miR-520e和AEG-1的靶向关系,通过qRT-PCR或Western blotting检测AEG-1、基质金属蛋白酶2（MMP2）、MMP9、NF-κB p65（p65）和磷酸化的NF-κB p65（p-p65）的表达。结果：与癌旁组织相比,结直肠癌组织中miR-520e的表达水平降低（t=9.353,P<0.001）。与对照组相比,miR-520e-mimic组的OD_490nm 值降低,细胞凋亡率升高,细胞迁移和侵袭数量降低,MMP2、MMP9和p-p65蛋白表达水平降低（P<0.001）。与对照组相比,miR-520e-inhibitor组的OD_490nm 值升高,细胞凋亡率降低,细胞迁移和侵袭数量升高,MMP2、MMP9和p-p65蛋白表达水平升高。与NC-mimic组相比,miR-520e-inhibitor组的相对荧光素酶活性降低（P<0.001）。与对照组相比,miR-520e-mimic组的AEG-1的mRNA和蛋白表达水平均降低,而miR-520e-inhibitor组均升高（P<0.001）。与miR-520e-mimic+NC-pcDNA3.1组相比,miR-520e-mimic+AEG-1-pcDNA3.1组的AEG-1的mRNA和蛋白表达水平升高,OD_490nm 值升高,细胞凋亡率降低,迁移和侵袭细胞数增加,MMP2和MMP9的蛋白表达水平及p65的磷酸化水平均增加（P<0.001）。结论：miR-520e在结直肠癌中表达降低,可通过靶向抑制AEG-1来发挥抗结直肠癌特性,其抗癌机制可能通过NF-κB信号通路介导。     

喉鳞状细胞癌组织miR-1207-5p、miR-186-5p表达水平与PI3K/Akt信号通路、临床病理特征和预后的关系

摘要 目的：探讨喉鳞状细胞癌（LSCC）组织miR-1207-5p、miR-186-5p表达水平与磷脂酰肌醇3-激酶/蛋白激酶B（PI3K/Akt）信号通路、临床病理特征和预后的关系。方法：选取2017年1月至2020年1月南充市中心医院收治的120例LSCC患者,取手术切除的LSCC组织和癌旁组织。检测miR-1207-5p、miR-186-5p、PI3K mRNA、Akt mRNA表达。患者出院后随访3年,统计总生存（OS）和无复发生存（RFS）情况。分析miR-1207-5p、miR-186-5p与PI3K、Akt的相关性以及影响LSCC患者预后的因素。结果：LSCC组织miR-1207-5p、miR-186-5p表达低于癌旁组织（P＜0.05）,PI3K mRNA、Akt mRNA表达高于癌旁组织（P＜0.05）。LSCC组织miR-1207-5p、miR-186-5p表达与PI3K mRNA、Akt mRNA表达呈负相关（P＜0.05）。肿瘤直径≥1 cm、低分化、TNM分期Ⅲ期、颈部淋巴结转移LSCC组织中miR-1207-5p、miR-186-5p表达低于肿瘤直径＜1 cm、中高分化、TNM分期Ⅰ～Ⅱ期、无颈部淋巴结转移（P＜0.05）。miR-1207-5p低表达、miR-186-5p低表达LSCC患者3年总生存（OS）率和无复发生存（RFS）率低于miR-1207-5p高表达、miR-186-5p高表达LSCC患者（P＜0.05）。多因素COX回归分析显示TNM III期、颈部淋巴结转移是LSCC患者复发和死亡的危险因素（P＜0.05）,高miR-1207-5p、高miR-186-5p是保护因素（P＜0.05）。结论：LSCC组织中miR-1207-5p和miR-186-5p表达均下调,与LSCC恶性病理特征、PI3K/Akt信号通路激活以及低生存率有关。     

miR-155-TP53INP1调节轴在结直肠癌化疗药物敏感性中的作用机制

摘要 目的：初步揭示miR-155通过靶向调节TP53INP1表达水平影响结直肠癌细胞对5-FU化疗敏感性。方法：将人结肠直肠癌细胞系HCT116进行培养,提取细胞总RNA后,采用miR-155逆转录特异性引物构建反转录体系进行PCR扩增,通过qRT-PCR检测miR-155在5-FU耐药细胞HCT116/FU及敏感细胞株HCT116中的表达情况;取对数生长期细胞,分别转染miR-155mimics、miR-155抑制剂、miR-155阴性对照后,采用CCK-8法检测miR-155对细胞5-FU药物敏感性的影响,双荧光素酶报告基因系统验证miR-155与TP53INP1的靶基因关系,Western blot检测miR-155对 TP53INP1表达的影响。结果：miR-155在HCT116 /Fu细胞中的表达量是HCT116细胞的7.25倍;在相同5-FU浓度时,HCT116+阴性对照的细胞生长抑制率均高于HCT116+mimics、半数抑制浓度显著低于HCT116+mimics,差异均具有统计学意义(P＜0.05);TP53INP1是miR-155的靶基因,能显著降低野生型TP53INP1 3''-UTR的荧光素酶活性;转染miR-155 mimics后,TP53INP1的相对表达量显著下降,转染miR-155抑制剂后,TP53INP1的相对表达量显著升高,差异均具有统计学意义(P＜0.05)。结论：miR-155水平升高使HCT116细胞对5-FU的敏感性降低,miR-155可能通过靶向调节TP53INP1的表达水平,从而影响结直肠癌细胞对5-FU的敏感性。     

肝细胞癌组织miR-124-3p、miR-212-5p表达与PI3K/Akt信号通路和预后的关系研究

摘要 目的：探讨肝细胞癌组织中微小核糖核酸（miR）-124-3p、miR-212-5p表达与磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）信号通路和预后的关系。方法：选择2017年9月至2019年9月徐州医科大学附属医院肝胆外科收治的93例肝细胞癌患者,采用实时荧光定量聚合酶链反应（qRT-PCR）检测肝细胞癌组织中miR-124-3p、miR-212-5p、PI3K 信使RNA（mRNA）、Akt mRNA的表达。Pearson相关性分析miR-124-3p、miR-212-5p表达与PI3K/Akt信号通路相关mRNA表达的相关性。绘制Kaplan-Meier生存曲线,Log-Rank检验不同miR-124-3p、miR-212-5p表达肝细胞癌患者3年总生存率（OS）的差异。结果：肝细胞癌组织中miR-124-3p表达低于癌旁组织（P＜0.05）,miR-212-5p、PI3K mRNA、Akt mRNA表达高于癌旁组织（P＜0.05）。肝细胞癌组织中miR-124-3p表达与PI3K mRNA、Akt mRNA表达呈负相关（P＜0.05）,miR-212-5p表达与PI3K mRNA、Akt mRNA表达呈正相关（P＜0.05）。Ⅱa期、低分化患者肝细胞癌组织中miR-124-3p表达低于Ⅰa-Ⅰb期、中高分化患者（P＜0.05）,miR-212-5p表达高于Ⅰa-Ⅰb期、中高分化患者（P＜0.05）。随访期间死亡37例, miR-124-3p低表达组3年OS为42.55%,低于miR-124-3p高表达组的78.26%（P＜0.05）,miR-212-5p高表达组3年OS为50.00%,低于miR-212-5p低表达组的71.11%（P＜0.05）。结论：肝细胞癌组织中miR-124-3p表达下调,miR-212-5p表达上调,且与肝细胞癌PI3K/Akt信号通路激活,PI3K mRNA和Akt mRNA高表达,低分化,CNLC分期Ⅱa期以及低OS有关。     

UPF1甲基化和miR-744-5p/CCND1在甲状腺乳头状癌中的作用机制研究

摘要 目的：探讨UPF1甲基化和miR-744-5p/CCND1在甲状腺乳头状癌中的作用机制研究。方法：将人甲状腺乳头状癌细胞株TCP-1和正常甲状腺上皮细胞Nthy-ori-3分别用去甲基化试剂5-Aza-CdR进行干预,分别在干预前后采用甲基化特异性PCR技术检测UPF1基因甲基化变化,采用Western-Blotting 检测干预UPF1、DNMT1、miR-744-5p、CCND1蛋白相对表达,采用transwell细胞侵袭实验检测细胞侵袭情况。结果：PCR扩增显示,UPF1基因在Nthy-ori-3组仅出现非甲基化引物扩增条带（U条带）,在TCP-1组仅出现甲基化引物扩增条带（M条带）。经5-Aza-Cdr作用后,UPF1基因甲基化扩增条带减少,甲基化表达降低。各组UPF1、DNMT1、miR-744-5p、CCND1蛋白相对表达差异具有统计学意义（P＜0.05）。与Nthy-ori-3组比较,TCP-1组DNMT1、UPF1蛋白相对表达明显提高,miR-744-5p、CCND1蛋白相对表达明显降低（P＜0.05）;与TCP-1组比较,TCP-1干预组DNMT1、UPF1蛋白相对表达明显降低,miR-744-5p、CCND1蛋白相对表达明显提高（P＜0.05）。与TCP-1组比较,TCP-1干预组细胞侵袭、迁移数量明显减少（P＜0.05）。结论：UPF1甲基化存在于甲状腺乳头状癌中,UPF1基因甲基化的表达缺失可能抑制miR-744-5p/CCND1轴,在甲状腺乳头状癌发生发展中发挥关键作用。     

下调lncRNA MCF2L-AS1靶向miR-33b-5p抑制胃癌细胞增殖、迁移、侵袭并促进凋亡

摘要 目的：探讨lncRNA MCF2L-AS1对胃癌细胞恶性生物学行为的影响及分子机制。方法：选取45例胃癌患者的癌组织及癌旁正常组织,或培养胃黏膜上皮细胞GES-1、胃癌细胞HGC-27,采用RT-qPCR检测MCF2L-AS1和miR-33b-5p的表达水平。采用双荧光素酶报告实验检测MCF2L-AS1和miR-33b-5p的靶向关系。将HGC-27细胞分为si-NC组、si-MCF2L-AS1组、mimic NC组、miR-33b-5p mimic组、si-MCF2L-AS1+inhibitor NC组、si-MCF2L-AS1+miR-33b-5p inhibitor组,分别转染si-NC、si-MCF2L-AS1、mimic NC、miR-33b-5p mimic或共转染si-MCF2L-AS1+inhibitor NC、si-MCF2L-AS1+miR-33b-5p inhibitor。采用MTT实验检测细胞增殖情况,流式细胞术检测细胞凋亡率,克隆形成实验检测细胞克隆形成数,Transwell实验检测迁移和侵袭细胞数。结果：与癌旁正常组织或GES-1细胞相比,胃癌组织或HGC-27细胞中MCF2L-AS1表达水平升高、miR-33b-5p表达水平降低,差异均有统计学意义（P<0.05）。MCF2L-AS1可靶向调控miR-33b-5p。下调MCF2L-AS1或过表达miR-33b-5p,miR-33b-5p表达水平升高,HGC-27细胞凋亡率升高,但细胞增殖、克隆形成数、迁移和侵袭数均减少,差异均有统计学意义（P<0.05）。抑制miR-33b-5p可减弱下调MCF2L-AS1对HGC-27细胞的生物学作用。结论：下调MCF2L-AS1通过上调miR-33b-5p抑制胃癌细胞增殖、迁移、侵袭并促进凋亡;MCF2L-AS1通过靶向调控miR-33b-5p表达进而参与胃癌细胞的恶性生物学行为。     

宫颈癌组织miR-200b-5p、miR-424-5p表达与临床病理特征、PI3K/AKT/mTOR信号通路和预后的关系研究

摘要 目的：探讨宫颈癌组织微小核糖核酸（miRNA）-200b-5p、miR-424-5p表达与临床病理特征、磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白（PI3K/AKT/mTOR）信号通路和预后的关系。方法：选取2016年7月～2019年6月西安市中心医院收治的123例宫颈癌患者,采用定量聚合酶链式反应（qPCR）检测癌组织与癌旁组织中miR-200b-5p、miR-424-5p、PI3K信使RNA（mRNA）、AKT mRNA、mTOR mRNA表达。分析miR-200b-5p、miR-424-5p表达与PI3K mRNA、AKT mRNA、mTOR mRNA表达的相关性及与临床病理特征的关系。采用K-M法绘制不同miR-200b-5p、miR-424-5p表达宫颈癌患者生存曲线。结果：与癌旁组织比较,宫颈癌组织中miR-200b-5p、miR-424-5p表达降低,PI3K mRNA、AKT mRNA、mTOR mRNA表达升高（P＜0.05）。Pearson相关性分析显示,宫颈癌组织中miR-200b-5p、miR-424-5p表达与PI3K mRNA、AKT mRNA、mTOR mRNA表达均呈负相关,PI3K mRNA、AKT mRNA、mTOR mRNA表达呈两两相关（P＜0.05）。miR-200b-5p、miR-424-5p表达与宫颈癌分化程度、国际妇产科联盟（FIGO）分期和淋巴结转移有关（P＜0.05）。随访3年,123例宫颈癌患者累积生存率为62.60%（77/123）。K-M生存曲线分析显示,miR-200b-5p、miR-424-5p高表达组累积生存率分别高于miR-200b-5p、miR-424-5p低表达组（P＜0.05）。结论：宫颈癌组织中miR-200b-5p、miR-424-5p低表达,与分化程度、FIGO分期、淋巴结转移、PI3K/AKT/mTOR信号通路和预后有关。     

miR-145、miR-186表达与非小细胞肺癌组织临床病理特征和PI3K/Akt/ mTOR信号通路的关系

摘要 目的：探讨微小核糖核酸（miR）-145、miR-186表达与非小细胞肺癌（NSCLC）临床病理特征和磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）/哺乳动物雷帕霉素靶蛋白（mTOR）信号通路的关系。方法：选择2020年3月至2023年3月我院收治的128例行肺癌根治手术治疗的NSCLC患者,取其术中癌组织和癌旁组织（距离癌组织5 cm以上）,采用实时荧光定量聚合酶链式反应（qRT-PCR）检测miR-145、miR-186以及PI3K mRNA、Akt mRNA、mTOR mRNA表达。分析miR-145、miR-186表达与NSCLC患者临床病理特征的关系;Pearson检验分析NSCLC患者癌组织miR-145、miR-186表达与PI3K mRNA、Akt mRNA、mTOR mRNA表达的相关性。结果：癌组织miR-145、miR-186表达低于癌旁组织（P＜0.05）,低分化、TNM ⅢA期、淋巴结转移的NSCLC组织miR-145、miR-186表达低于中高分化、TNM Ⅰ～Ⅱ期、未发生淋巴结转移的NSCLC组织（P＜0.05）。癌组织PI3K mRNA、Akt mRNA 、mTOR mRNA表达均高于癌旁组织（P＜0.05）,癌组织miR-145、miR-186表达与PI3K mRNA、Akt mRNA、mTOR mRNA表达均呈负相关（P＜0.05）。结论：NSCLC组织中miR-145、miR-186表达下调,miR-145、miR-186表达下调可能激活PI3K/ Akt/mTOR信号通路促使NSCLC进展,且与NSCLC患者癌组织低分化、TNM ⅢA期、淋巴结转移有关。     

上皮性卵巢癌组织miR-338-3p、miR-1294的表达及临床意义

摘要 目的：探讨上皮性卵巢癌组织中微小RNA（miR）-338-3p、miR-1294表达与患者临床病理参数的关系,并分析其表达对上皮性卵巢癌预后的影响。方法：收集2010年6月～2015年6月我院行手术治疗的上皮性卵巢癌患者的石蜡组织样本（癌组织和癌旁正常组织）,实时荧光定量PCR（RT-PCR）技术检测组织样本中miR-338-3p、miR-1294的表达情况;分析癌组织中miR-338-3p、miR-1294表达与患者临床病理参数的关系;Kaplan-Meier生存曲线分析miR-338-3p、miR-1294表达与患者预后的关系;Cox回归模型分析上皮性卵巢癌患者的预后影响因素。结果：与癌旁正常组织相比,上皮性卵巢癌组织中miR-338-3p、miR-1294表达均降低（均P<0.05）;miR-338-3p表达与组织学分级、淋巴结转移、FIGO分期相关（均P<0.05）;miR-1294表达与肿瘤直径、淋巴结转移、FIGO分期相关（均P<0.05）。Kaplan-Meier生存曲线结果显示,miR-338-3p低表达组患者、miR-1294低表达组患者的预后较差（均P<0.05）。Cox回归模型分析结果显示,淋巴结转移、较高FIGO分期、miR-338-3p低表达、miR-1294低表达是上皮性卵巢癌预后的危险因素（均P<0.05）。结论：上皮性卵巢癌组织中miR-338-3p、miR-1294均低表达,且均与较差临床病理参数、预后不良相关;miR-338-3p、miR-1294可能是上皮性卵巢癌患者预后预测的潜在指标。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.