During thyroidectomy, inadvertent parathyroid damage commonly occurs owing to the varying location of the parathyroid glands and their similarity to adipose tissue, lymph nodes, and thyroid nodules. Therefore, in this study, we aimed to develop a novel self-assembled nanobody-DNA nanomachine conjugate probe (PTH-Nb-HCR) for precisely identifying the parathyroid glands and monitoring parathyroid hormone (PTH) levels in patient blood. The probe consists of two components: the first component is the nanobody (Nb)-nucleic acid conjugate PTH-Nb-Trigger, acting as a recognition unit for PTH and capable of initiating the hybridization chain reaction (HCR), whereas the second component comprises the HCR hairpin probes HP1-FAM (with a modified FAM signal molecule at the 5′ end) and HP2. The assembly of the PTH-Nb-HCR fluorescent probe takes only 20 min. Using the probe, the serum PTH level was quantitatively detected in the 0.1–1000 pg/mL range, and the parathyroid glands were rapidly distinguished from adipose, lymphoid, and thyroid tissues within 5 min. We also used this probe for imaging parathyroid glands with hyperparathyroidism. Considering the strong correlation between fluorescence intensity and preoperative serum PTH levels in patients, we anticipate the use of this probe to distinguish normal and hyperfunctional tissues during hyperparathyroidism surgery.

中文翻译：

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自组装纳米抗体-DNA纳米机器缀合物用于甲状旁腺术中成像和甲状旁腺激素检测

在甲状腺切除术中，由于甲状旁腺的位置不同且与脂肪组织、淋巴结和甲状腺结节相似，因此通常会发生无意的甲状旁腺损伤。因此，在本研究中，我们旨在开发一种新型自组装纳米抗体-DNA纳米机器结合探针（PTH-Nb-HCR），用于精确识别甲状旁腺并监测患者血液中的甲状旁腺激素（PTH）水平。该探针由两个组件组成：第一个组件是纳米抗体（Nb）-核酸缀合物PTH-Nb-Trigger，作为PTH的识别单元并能够启动杂交链式反应（HCR），而第二个组件包括HCR 发夹探针 HP1-FAM（5' 端带有修饰的 FAM 信号分子）和 HP2。 PTH-Nb-HCR 荧光探针的组装仅需 20 分钟。使用该探针可在0.1-1000 pg/mL范围内定量检测血清PTH水平，并在5分钟内快速区分甲状旁腺与脂肪、淋巴和甲状腺组织。我们还使用该探头对患有甲状旁腺功能亢进的甲状旁腺进行成像。考虑到荧光强度与患者术前血清 PTH 水平之间的强相关性，我们预计在甲状旁腺功能亢进手术期间使用该探针来区分正常组织和功能亢进组织。