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Infrared vibrational spectroscopy: a rapid and novel diagnostic and monitoring tool for cystinuria

机译:红外振动光谱:一种快速,新颖的胱氨酸尿症诊断和监测工具

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摘要

Cystinuria is the commonest inherited cause of nephrolithiasis (~1% in adults; ~6% in children) and is the result of impaired cystine reabsorption in the renal proximal tubule. Cystine is poorly soluble in urine with a solubility of ~1 mM and can readily form microcrystals that lead to cystine stone formation, especially at low urine pH. Diagnosis of cystinuria is made typically by ion-exchange chromatography (IEC) detection and quantitation, which is slow, laboursome and costly. More rapid and frequent monitoring of urinary cystine concentration would significantly improve the diagnosis and clinical management of cystinuria. We used attenuated total reflection - Fourier transform infrared spectroscopy (ATR-FTIR) to detect and quantitate insoluble cystine in 22 cystinuric and 5 healthy control urine samples. Creatinine concentration was also determined by ATR-FTIR to adjust for urinary concentration/dilution. Urine was centrifuged, the insoluble fraction re-suspended in 5 μL water and dried on the ATR prism. Cystine was quantitated using its 1296 cm−1 absorption band and levels matched with parallel measurements made using IEC. ATR-FTIR afforded a rapid and inexpensive method of detecting and quantitating insoluble urinary cystine. This proof-of-concept study provides a basis for developing a high-throughput, cost-effective diagnostic method for cystinuria, and for point-of-care clinical monitoring.
机译:半胱氨酸尿症是肾结石病的最常见遗传原因(成人约1%;儿童约6%),并且是肾近端小管中胱氨酸重吸收受损的结果。胱氨酸在尿液中的溶解度很低,溶解度约为1MmM,并且容易形成微晶,导致胱氨酸结石的形成,特别是在尿液pH值较低的情况下。胱氨酸尿症的诊断通常是通过离子交换色谱法(IEC)检测和定量来进行的,该过程缓慢,费力且昂贵。更快,更频繁地监测尿中胱氨酸的浓度将显着改善胱氨酸尿症的诊断和临床管理。我们使用了衰减全反射-傅立叶变换红外光谱(ATR-FTIR)来检测和定量22个胱氨酸尿酸和5个健康对照尿液样品中的不溶性胱氨酸。还可以通过ATR-FTIR测定肌酐浓度,以调整尿液浓度/稀释度。离心尿液,将不溶级分重悬于5μL水中,并在ATR棱镜上干燥。胱氨酸使用其1296 cm-1吸收带进行定量,其水平与使用IEC进行的平行测量相匹配。 ATR-FTIR提供了一种快速且廉价的检测和定量不溶性尿胱氨酸的方法。这项概念验证研究为开发高通量,具有成本效益的胱氨酸尿症诊断方法以及即时医疗临床监测提供了基础。

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