Comparison of RT-PCR, RT-nested PCRs, and real-time PCR for diagnosis of severe fever with thrombocytopenia syndrome: a prospective study

Abstract

We designed a highly sensitive reverse transcription nested polymerase chain reaction targeting the M-segment (NPCR-M) of severe fever with thrombocytopenia syndrome (SFTS) virus. NPCR-M was performed in parallel with three other referenced PCR assays QPCR-S, PCR-M, and NPCR-S to assess their clinical usefulness as routine diagnostic techniques for SFTS. In this multi-centered prospective study, 122 blood samples from 38 laboratory-confirmed SFTS patients and 85 control samples were used. The results demonstrated that QPCR-S and NPCR-S had better sensitivity rate up to 21 days after symptom onset however, the PCR-M showed poor sensitivity after 7 days of symptom onset. Our designed NPCR-M had a higher detection rate up to 40 days from symptom onset and revealed the persistence of SFTSV RNA in the early convalescent phase. No false-positive results were seen for the control samples. Additionally, NPCR-M showed positive results for a sample that initially showed negative results from other PCRs and for many other samples collected in the convalescent phase of SFTS. Our designed nested PCR is suitable for SFTSV detection in patients’ blood collected in the acute and early convalescent phase of SFTS, and shows better sensitivity and high specificity even up to 40 days after symptom onset.

Accuracy of reverse‐transcription polymerase chain reaction and loop‐mediated isothermal amplification in diagnosing severe fever with thrombocytopenia syndrome: A systematic review and meta‐analysis

“…However, currently, conventional RT-PCR, nested RT-PCR, and real-time RT-PCR are used for the detection of SFTS. 19 In our study, we found heterogeneity in RT-PCR caused by threshold effects. This may be because of the different cycle threshold (Ct) values used as criteria for determining positivity in the studies analyzed.…”

“…Additionally, researchers can also develop new and more effective primers/probes for PCR research and develop more precise commercial kits for the diagnosis of SFTS. However, currently, conventional RT‐PCR, nested RT‐PCR, and real‐time RT‐PCR are used for the detection of SFTS 19 …”

“…18 Meanwhile, ELISA and other traditional virus detection techniques have some disadvantages; they are timeconsuming, expensive, and complicated to perform. 4,19 Nevertheless, nucleic acid detection diagnostic methods are rapid, quantitative, and highly sensitive. 4 Reverse-transcription polymerase chain reaction (RT-PCR) can detect low-copy viral RNA quantitatively and is used for the early diagnosis of SFTS when viral immunoglobulin M/G cannot be detected.…”

“…With the progress of technology, significant developments have been likewise made in nucleic acid molecular diagnosis testing, expanding its applications to various scenarios and detection targets 15–17 and turning it into the gold standard of many virological diseases, such as coronavirus disease 2019, an acute infectious disease that is spreading globally 18 . Meanwhile, ELISA and other traditional virus detection techniques have some disadvantages; they are time‐consuming, expensive, and complicated to perform 4,19 . Nevertheless, nucleic acid detection diagnostic methods are rapid, quantitative, and highly sensitive 4 …”

“… 18 Meanwhile, ELISA and other traditional virus detection techniques have some disadvantages; they are time‐consuming, expensive, and complicated to perform. 4 , 19 Nevertheless, nucleic acid detection diagnostic methods are rapid, quantitative, and highly sensitive. 4 …”