Application of miR-376b and detection kit
A technology of mir-376b and 1.mir-376b, which is applied in the field of acute kidney injury detection in patients with sepsis, can solve the problems of lack of specificity in diagnosis, and achieve the effects of high diagnostic sensitivity, non-degradation and mortality reduction.
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Embodiment 1
[0040] Gene chip analysis: AKI induced by intraperitoneal injection of lipopolysaccharide (LPS) in mice with sepsis, and microRNA expression profile changes in kidney tissue were analyzed by chip technology. The results showed that after 24 hours of LPS treatment, the expression of some microRNAs was up-regulated or down-regulated, including miR-376b. These results reveal the possible changing trend of microRNA in sepsis AKI and lay the foundation for the next experiments.
[0041] Animal experiment design:
[0042] C57BL / 6 mice, 6-8 weeks old, were purchased from Changsha Slake Jingda Experimental Animal Co., Ltd. We induced sepsis animal model by intraperitoneal injection (dose 10 mg / kg) of LPS (purchased from Sigma, USA). The specific grouping is shown in Table 1: (LPS was dissolved in normal saline, and the control group was simply injected with normal saline)
[0043] Table 1
[0044]
[0045] After the injection, the mice were put back into the breeding cage and p...
Embodiment 2
[0050] In addition to animal experiments, clinical specimens were collected at the same time, and relevant clinical tests and tests were carried out. A total of 40 urine samples from sepsis patients in our hospital (the Second Xiangya Hospital of Central South University) were collected; 20 cases of sepsis complicated with AKI, 20 cases of sepsis without AKI (control) (all sepsis cases The patients were previously healthy when they were collected, and some basic indicators and inflammatory indicators are shown in Table 2 below).
[0051] Table 2
[0052] sepsis non-AKI sepsis-AKI P value Gender: Male Female) 13 / 7 8 / 12 0.12 age 54.85±4.11 55.20±4.13 0.94 Serum creatinine, μmol / L 76.71±7.25 345.20±53.63 <0.01*
[0053] Total RNA was extracted from the patient's urine sample, cDNA was synthesized using taqman probe (human-derived miR-376b probe, primer catalog number: 001102, taqman microRNA Reverse Transcirculation Kit, all purchased from ...
Embodiment 3
[0055] We used receiver operating characteristic curve (ROC) to evaluate the validity of urinary miR-376b as a diagnostic indicator of sepsis AKI. The result is as Figure 6 , AUC=0.81, which has good diagnostic value. According to the ROC curve, we believe that the diagnostic effect is the best when the Youden index (Youden index=sensitivity+specificity-1) is maximum, and the diagnostic cut-off value determined according to this principle is urinary miR-376b<0.69 times. The final diagnostic cut-off values and their diagnostic sensitivity and specificity are shown in Table 3.
[0056] table 3
[0057]
[0058] Urinary [TIMP2]*[IGFBP7] is a sensitive indicator approved by the US FDA for the early diagnosis of AKI, and its diagnostic value in AKI has been widely recognized. The present invention uses the same sample to measure urine [TIMP2]*[IGFBP7], and the ROC curve of urine [TIMP2]*[IGFBP7] to diagnose sepsis AKI is shown in Figure 7 , indicating that compared with ...
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