Khan Mohammed Faraaz, Roopa C
Department of Microbiology, Kamineni Institute of Medical Sciences, Narketpally, Telangana, India.
Department of Microbiology, SVS Medical College, Mahabubnagar, Telangana, India.
Can J Infect Dis Med Microbiol. 2023 Aug 23;2023:6624932. doi: 10.1155/2023/6624932. eCollection 2023.
Conventional nucleic acid extraction involves usage of spin columns to isolate the RNA, but this is labor intensive. This study compares the spin column method with a dry swab-based method of extraction using a proteinase K buffer and subsequent heat inactivation. A total of 56 subjects were tested for COVID-19 by RT-PCR with probes targeting the E and RdRp genes by collecting two nasopharyngeal and two oropharyngeal swabs and subjecting one set to nucleic acid extraction by spin column and the other set to dry swab-based methods. Out of the 56 samples tested, 27 were positive for VTM-based extraction and 29 were negative. Dry swab-based extraction produced 22 positive results (sensitivity = 81.48%) and 34 negative results. The E gene was detectable in 25 samples by the dry swab method out of 27 samples that tested positive by the VTM-based method (sensitivity = 92.5%). The RdRp gene was detectable in 22 samples by the dry swab method out of 27 samples that tested positive by the VTM-based method (sensitivity = 81.48%). Concordance was 91% with discordance at 9% and a Kappa value of 0.82, indicating almost perfect agreement between the two methods. Our findings indicate that the dry swab method of nucleic acid extraction is a useful alternative to conventional spin column-based extraction with comparable sensitivity and specificity. The trial was registered with the Clinical Trials Registry of India (CTRI) with a CTRI registration number of CTRI/2021/12/038792.
传统的核酸提取需要使用离心柱来分离RNA,但这一过程 labor intensive。本研究将离心柱法与一种基于干拭子的提取方法进行了比较,该方法使用蛋白酶K缓冲液并随后进行热灭活。通过收集两根鼻咽拭子和两根口咽拭子,将其中一组样本用离心柱法进行核酸提取,另一组样本用基于干拭子的方法进行核酸提取,共有56名受试者通过针对E基因和RdRp基因的探针进行RT-PCR检测COVID-19。在检测的56个样本中,基于病毒运输培养基(VTM)的提取方法有27个呈阳性,29个呈阴性。基于干拭子的提取方法产生了22个阳性结果(灵敏度 = 81.48%)和34个阴性结果。在基于VTM的方法检测呈阳性的27个样本中(灵敏度 = 92.5%),通过干拭子法在25个样本中检测到了E基因。在基于VTM的方法检测呈阳性的27个样本中(灵敏度 = 81.48%),通过干拭子法在22个样本中检测到了RdRp基因。一致性为91%,不一致性为9%,Kappa值为0.82,表明两种方法之间几乎完全一致。我们的研究结果表明,核酸提取干拭子法是传统离心柱法的一种有用替代方法,具有相当的灵敏度和特异性。该试验已在印度临床试验注册中心(CTRI)注册,CTRI注册号为CTRI/2021/12/038792。