高光熱效率之聚吡咯包覆銅摻雜普魯士藍結合具可變光源之攜帶式光熱裝置應用於多重疾病核酸檢測

Abstract

當前聚合酶鏈式反應 (Polymerase chain reaction, PCR) 做為呼吸道病毒的黃金檢測標準，受限於熱循環之升降溫速度，導致步驟耗時，因此僅能提供有限的檢測量能。為了解決此問題，本研究開發出一種高光熱轉換效率之聚吡咯包覆銅摻雜普魯士藍奈米方塊 (polypyrrole-coated copper-doped Prussian blue Nanocubes, PPy-CuPB)，並結合以調控頻率來改變亮度的光熱 PCR 系統。材料藉由銅摻雜來提升鍵結強度、電子導電性與能帶邊緣調控能力，之後包覆具有剛性π共軛結構之聚吡咯，顯著提升穩定性與紅外光吸收。此外，搭配脈衝寬度調變 (pulse-width modulation, PWM) 控制之光源，提供不同溫度條件下光強度控制，提升溫控穩定性。材料經密度泛函理論模擬與實測顯示，銅摻雜會引發帶隙變窄效應與增強非輻射復合，最終包覆聚吡咯後可使光熱轉換效率提高至 52.1 %，實現在 740 nm 光源下快速升溫 (11.75 °C/s) 與 30 次穩定熱循環。同時本研究成功擴展光熱 PCR 至多重聚合酶鏈式反應 (Multiplex PCR)，成功擴增嚴重急性呼吸道症候群冠狀病毒2型 (Severe Acute Respiratory Syndrome Coronavirus 2, SARS-CoV-2) Omicron 與 A 型流感病毒 (influenza A virus, IAV) H3N2 之雙重標的，標準品之檢測極限分別達 0.6 pg/mL 與 14.87 pg/mL。最後於真實臨床檢體中驗證本系統之接收者操作特徵 (receiver operating characteristic, ROC) 曲線之曲線下面積 (Area under the Curve, AUC) 達 0.98 與 1.00。吾人所開發之平台整合光熱奈米材料與可調控光源裝置，具備應用於快速且多重之疾病診斷的潛力，未來可做為臨床檢測與公共衛生監控的技術基礎。

Polymerase chain reaction (PCR) is the gold standard for detecting respiratory viruses. However, the process of PCR is time-consuming due to limitations in heating and cooling efficiency, as well as complex thermal cycling steps. To solve this problem, this research developed a novel high photothermal conversion efficiency nanocomposite polypyrrole-coated copper-doped Prussian blue Nanocubes (PPy-CuPB) and combined it with a photothermal PCR device, which can control light intensity by adjusting power frequency. First, the core material, Prussian blue, was doped with copper to enhance bond strength, electrical conductivity and band edge control. Second, it was coated with polypyrrole, which has a rigid π-conjugated structure, and it significantly upgraded infrared absorption and dispersibility. Finally, this study produced a pulse-width modulation (PWM) controlled light source, which enables adjustment of light intensity under different temperatures and improves temperature stability during reactions. The density functional theory calculations and experimental results further revealed that copper doping causes bandgap narrowing and raises nonradiative recombination. After copper doping and polypyrrole coating, the photothermal conversion efficiency of PPy-CuPB reached 52.1%. As a result, it enabled rapid heating at 11.75 °C/s under 740 nm irradiation and maintained stable thermal cycling over 30 cycles. Eventually, we successfully extended the photothermal PCR to multiplex targets, such as SARS-CoV-2 Omicron and influenza A virus (IAV) H3N2. The results demonstrated limits of detection of 0.6 pg/mL for SARS-CoV-2 and 14.87 pg/mL for IAV using standard samples. Moreover, receiver operating characteristic (ROC) curve analysis produced an Area under the Curve (AUC) of 0.99 for SARS-CoV-2 and 1.00 for IAV based on real clinical samples. This platform, which integrates nanomaterials with a tunable light source, held significant potential for rapid and multiplex disease diagnostics. In the future, it could serve as a foundation for clinical testing and public health surveillance.