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Discrimination of False Negative Results in RT-PCR Detection of SARS-CoV-2 RNAs in Clinical Specimens by Using an internal reference

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Virologica Sinica 2020年第6期35卷 758-767页

作者： Yafei Zhang Changtai Wang Mingfeng Han Jun Ye Yong Gao Zhongping Liu Tengfei He Tuantuan Li Mengyuan Xu Luping Zhou Guizhou Zou Mengji Lu Zhenhua Zhang Department of Infectious Diseases The Second Hospital of Anhui Medical UniversityHefei 230601China Institute of Clinical Virology The Second Hospital of Anhui Medical UniversityHefei 230601China Department of internal Medicine The Second Hospital of FuyangFuyang 236015China Institute for Virology University Hospital of EssenUniversity of Duisburg-Essen45147 EssenGermany

Reverse transcription-polymerase chain reaction(RT-PCR)is an essential method for specific diagnosis of SARS-CoV-2 ***,false negative test results are often *** this study,we attempted to determine the principal causes leading to false negative results of RT-PCR detection of SARS-CoV-2 RNAs in respiratory tract *** sputum and throat swab specimens from 161 confirmed COVID-19 patients were tested with a commercialfluorescent RT-PCR kit targeting the ORF1 ab and N regions of SARS-CoV-2 *** RNA level of a cellular housekeeping gene ribonuclease P/MRP subunit p30(RPP30)in these specimens was also assessed by *** for a total of 1052 samples were retrospectively re-analyzed and a strong association between positive results in SARS-CoV-2 RNA tests and high level of RPP30 RNA in respiratory tract specimens was *** using the ROC-AUC analysis,we identified Ct cutoff values for RPP30 RT-PCR which predicted false negative results for SARS-CoV-2 RT-PCR with high sensitivity(95.03%–95.26%)and specificity(83.72%–98.55%)for respective combination of specimen type and ampli-fication *** these Ct cutoff values,false negative results could be reliably ***,the presence of cellular materials,likely infected host cells,are essential for correct SARS-CoV-2 RNA detection by RT-PCR in patient ***30 could serve as an indicator for cellular content,or a surrogate indicator for specimen *** addition,our results demonstrated that false negativity accounted for a vast majority of contradicting results in SARS-CoV-2 RNA test by RT-PCR.

关键词： COVID-19 SARS-CoV-2 RT-PCR False negative results internal reference

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Multiplex ratiometric gold nanoprobes based on surface-enhanced Raman scattering enable accurate molecular detection and imaging of bladder cancer

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Nano Research 2022年第4期15卷 3487-3495页

作者： Xiao Liang Pu Zhang Minghai Ma Tao Yang Xiangwei Zhao Rui Zhang Minxuan Jing Rundong Song Lei Wang Jinhai Fan Department of Urology The First Affiliated HospitalXi’an Jiaotong UniversityXi’an 710061China Department of Thoracic Surgery Tangdu HospitalAir Force Medical UniversityXi’an 710038China Department of Urology Zhejiang Provincial People’s HospitalPeople’s Hospital of Hangzhou Medical CollegeHangzhou 310014China State Key Laboratory of Bioelectronics School of Biological Science and Medical EngineeringNational Demonstration Center for Experimental Biomedical Engineering EducationSoutheast UniversityNanjing 210096China

Recently,surface-enhanced Raman scattering(SERS)has been successfully used in the non-invasive detection of bladder tumor(BCa).The internal standard method was considered as an effective ratiometric strategy for calibrating signal fluctuation originated from the interference of measurement conditions and ***,it is still difficult to detect the target mRNA quantitatively using the current ratiometric SERS *** this study,we developed an internal reference based ratiometric SERS *** kinds of molecular beacons(MB)carrying Raman reporter molecules were anchored on sea-urchinlike Au nanoclusters(AuNCs).Thymidine kinase1(TK1)MBs with hexachlorofluorescein(HEX)were used to capture tumor biomarker TK1 mRNA,and glyceraldehyde 3-phosphate dehydrogenase(GAPDH)MBs with 5(6)-carboxyfluorescein(FAM)were used to offer internal standard *** internal reference GAPDH MB can reflect the consistent content of the GAPDH mRNA in single *** ratiometric method(I745/I645)can more accurately reflect the content of target mRNA in single *** ratiometric nanoprobes had excellent stability(coefficient of variation:0.3%),high sensitivity(detection limit:3.4 pM),high specificity(capable of single-base mismatch recognition)and ribozyme-resistant ***,the nanoprobes can effectively distinguish BCa cells from normal cells,and it was easy to contour the single BCa cell using the ratiometric *** combining asymmetric polymerase chain reaction(PCR)and ratiometric nanoprobes,it was easy to distinguish the SERS ratio(I_(745)/I_(645))as low concentration as 10-14 *** clinical detection in urine samples from patients with BCa confirmed its potential for early noninvasive diagnosis of BCa with the sensitivity of 80%and specificity of 100%,which is superior to the current urine cytological method.

关键词： surface-enhanced Raman scattering(SERS) internal reference bladder cancer non-invasive detection SERS imaging

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