Development of an in-house dual RT-qPCR assay for detecting SFTSV and Hantaan virus simultaneously
Given the overlapping endemic regions and clinical similarities between severe fever with thrombocytopenia syndrome (SFTS) and hemorrhagic fever with renal syndrome (HFRS), we developed a dual real‐time fluorescence‐based reverse transcription quantitative polymerase chain reaction (RT-qPCR) method....
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-04-01
|
| Series: | Biosafety and Health |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590053625000436 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849711885261209600 |
|---|---|
| author | Xiaoyu Xue Youde Liu Chuan Song Tingting Liu Zishuai Liu Wenjing Niu Zhouling Jiang Yanli Xu Yuanyuan Zhang Ling Lin Zhihai Chen |
| author_facet | Xiaoyu Xue Youde Liu Chuan Song Tingting Liu Zishuai Liu Wenjing Niu Zhouling Jiang Yanli Xu Yuanyuan Zhang Ling Lin Zhihai Chen |
| author_sort | Xiaoyu Xue |
| collection | DOAJ |
| description | Given the overlapping endemic regions and clinical similarities between severe fever with thrombocytopenia syndrome (SFTS) and hemorrhagic fever with renal syndrome (HFRS), we developed a dual real‐time fluorescence‐based reverse transcription quantitative polymerase chain reaction (RT-qPCR) method. Recombinant plasmids and synthetic ribonucleic acid (RNA) were constructed to evaluate the specificity, sensitivity and reproducibility of the assay. Additionally, we assessed the specificity of the assay using samples from three distinct groups: individuals with confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection (n = 10), influenza A-positive individuals (n = 10), and healthy controls. Receiver operating characteristic (ROC) curves were used to assess diagnostic accuracy, while the Kappa coefficient and linear regression analysis were employed to evaluate clinical applicability. Our method exhibited specificity for both SFTSV and Hantaan virus detection, with detection limits of 333 and 1,022 copies/mL using plasmids, and 1,247 and 898 copies/mL using synthetic RNA, respectively. We evaluated 100 clinical samples from each of SFTS and HFRS. The Kappa coefficients for both diseases were 0.96. The areas under the ROC curves were 0.991 (P < 0.001) and 0.989 (P < 0.001), respectively. The linear regression equations were as follows: log (y) = 0.19 + 0.99 log (x) (R2 = 0.95) for SFTS virus, and log (y) = 0.01 + 0.65 log (x) (R2 = 0.92) for Hantaan virus. We established an in-house RT-qPCR method for the rapid quantification of both pathogens, making it an ideal tool for early clinical differentiation. |
| format | Article |
| id | doaj-art-c9346e89be9d4512933e9c65dfed1f15 |
| institution | DOAJ |
| issn | 2590-0536 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biosafety and Health |
| spelling | doaj-art-c9346e89be9d4512933e9c65dfed1f152025-08-20T03:14:30ZengElsevierBiosafety and Health2590-05362025-04-017211011610.1016/j.bsheal.2025.03.007Development of an in-house dual RT-qPCR assay for detecting SFTSV and Hantaan virus simultaneouslyXiaoyu Xue0Youde Liu1Chuan Song2Tingting Liu3Zishuai Liu4Wenjing Niu5Zhouling Jiang6Yanli Xu7Yuanyuan Zhang8Ling Lin9Zhihai Chen10Department of Infectious Disease, Peking University Ditan Teaching Hospital, Beijing 100015, ChinaDepartment of Infectious Diseases, Yantai Qishan Hospital, Yantai 264001, ChinaNational Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China; Beijing Key Laboratory of Emerging Infectious Diseases, Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China; Beijing Institute of Infectious Diseases, Beijing 100015, China; National Center for Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, ChinaNational Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China; Beijing Key Laboratory of Emerging Infectious Diseases, Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China; Beijing Institute of Infectious Diseases, Beijing 100015, China; National Center for Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, ChinaNational Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China; Department of Infectious Disease, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, ChinaNational Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China; Department of Infectious Disease, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, ChinaNational Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China; Department of Infectious Disease, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, ChinaDepartment of Infectious Diseases, Yantai Qishan Hospital, Yantai 264001, ChinaNational Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China; Beijing Key Laboratory of Emerging Infectious Diseases, Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China; Beijing Institute of Infectious Diseases, Beijing 100015, China; National Center for Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China; Corresponding authors: Capital Medical University, 8 Jingshun East Street, Chaoyang District, Beijing, China (Y. Zhang and Z. Chen); Yantai Qishan Hospital, 62 Huanshan Road, Zhifu District, Yantai City, Shandong Province, China (L. Lin).Department of Infectious Diseases, Yantai Qishan Hospital, Yantai 264001, China; Corresponding authors: Capital Medical University, 8 Jingshun East Street, Chaoyang District, Beijing, China (Y. Zhang and Z. Chen); Yantai Qishan Hospital, 62 Huanshan Road, Zhifu District, Yantai City, Shandong Province, China (L. Lin).Department of Infectious Disease, Peking University Ditan Teaching Hospital, Beijing 100015, China; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China; Department of Infectious Disease, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China; Beijing Research Center for Respiratory Infectious Diseases, Beijing 100015, China; Corresponding authors: Capital Medical University, 8 Jingshun East Street, Chaoyang District, Beijing, China (Y. Zhang and Z. Chen); Yantai Qishan Hospital, 62 Huanshan Road, Zhifu District, Yantai City, Shandong Province, China (L. Lin).Given the overlapping endemic regions and clinical similarities between severe fever with thrombocytopenia syndrome (SFTS) and hemorrhagic fever with renal syndrome (HFRS), we developed a dual real‐time fluorescence‐based reverse transcription quantitative polymerase chain reaction (RT-qPCR) method. Recombinant plasmids and synthetic ribonucleic acid (RNA) were constructed to evaluate the specificity, sensitivity and reproducibility of the assay. Additionally, we assessed the specificity of the assay using samples from three distinct groups: individuals with confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection (n = 10), influenza A-positive individuals (n = 10), and healthy controls. Receiver operating characteristic (ROC) curves were used to assess diagnostic accuracy, while the Kappa coefficient and linear regression analysis were employed to evaluate clinical applicability. Our method exhibited specificity for both SFTSV and Hantaan virus detection, with detection limits of 333 and 1,022 copies/mL using plasmids, and 1,247 and 898 copies/mL using synthetic RNA, respectively. We evaluated 100 clinical samples from each of SFTS and HFRS. The Kappa coefficients for both diseases were 0.96. The areas under the ROC curves were 0.991 (P < 0.001) and 0.989 (P < 0.001), respectively. The linear regression equations were as follows: log (y) = 0.19 + 0.99 log (x) (R2 = 0.95) for SFTS virus, and log (y) = 0.01 + 0.65 log (x) (R2 = 0.92) for Hantaan virus. We established an in-house RT-qPCR method for the rapid quantification of both pathogens, making it an ideal tool for early clinical differentiation.http://www.sciencedirect.com/science/article/pii/S2590053625000436Severe fever with thrombocytopenia syndrome virus (SFTSV)Hantaan virusHemorrhagic fever with renal syndrome (HFRS)Severe fever with thrombocytopenia syndrome (SFTS)Differential diagnosisDual real time reverse transcription quantitative polymerase chain reaction (RT-qPCR) |
| spellingShingle | Xiaoyu Xue Youde Liu Chuan Song Tingting Liu Zishuai Liu Wenjing Niu Zhouling Jiang Yanli Xu Yuanyuan Zhang Ling Lin Zhihai Chen Development of an in-house dual RT-qPCR assay for detecting SFTSV and Hantaan virus simultaneously Biosafety and Health Severe fever with thrombocytopenia syndrome virus (SFTSV) Hantaan virus Hemorrhagic fever with renal syndrome (HFRS) Severe fever with thrombocytopenia syndrome (SFTS) Differential diagnosis Dual real time reverse transcription quantitative polymerase chain reaction (RT-qPCR) |
| title | Development of an in-house dual RT-qPCR assay for detecting SFTSV and Hantaan virus simultaneously |
| title_full | Development of an in-house dual RT-qPCR assay for detecting SFTSV and Hantaan virus simultaneously |
| title_fullStr | Development of an in-house dual RT-qPCR assay for detecting SFTSV and Hantaan virus simultaneously |
| title_full_unstemmed | Development of an in-house dual RT-qPCR assay for detecting SFTSV and Hantaan virus simultaneously |
| title_short | Development of an in-house dual RT-qPCR assay for detecting SFTSV and Hantaan virus simultaneously |
| title_sort | development of an in house dual rt qpcr assay for detecting sftsv and hantaan virus simultaneously |
| topic | Severe fever with thrombocytopenia syndrome virus (SFTSV) Hantaan virus Hemorrhagic fever with renal syndrome (HFRS) Severe fever with thrombocytopenia syndrome (SFTS) Differential diagnosis Dual real time reverse transcription quantitative polymerase chain reaction (RT-qPCR) |
| url | http://www.sciencedirect.com/science/article/pii/S2590053625000436 |
| work_keys_str_mv | AT xiaoyuxue developmentofaninhousedualrtqpcrassayfordetectingsftsvandhantaanvirussimultaneously AT youdeliu developmentofaninhousedualrtqpcrassayfordetectingsftsvandhantaanvirussimultaneously AT chuansong developmentofaninhousedualrtqpcrassayfordetectingsftsvandhantaanvirussimultaneously AT tingtingliu developmentofaninhousedualrtqpcrassayfordetectingsftsvandhantaanvirussimultaneously AT zishuailiu developmentofaninhousedualrtqpcrassayfordetectingsftsvandhantaanvirussimultaneously AT wenjingniu developmentofaninhousedualrtqpcrassayfordetectingsftsvandhantaanvirussimultaneously AT zhoulingjiang developmentofaninhousedualrtqpcrassayfordetectingsftsvandhantaanvirussimultaneously AT yanlixu developmentofaninhousedualrtqpcrassayfordetectingsftsvandhantaanvirussimultaneously AT yuanyuanzhang developmentofaninhousedualrtqpcrassayfordetectingsftsvandhantaanvirussimultaneously AT linglin developmentofaninhousedualrtqpcrassayfordetectingsftsvandhantaanvirussimultaneously AT zhihaichen developmentofaninhousedualrtqpcrassayfordetectingsftsvandhantaanvirussimultaneously |