Matrix metallopeptidase 9 (MMP9) was found to be associated with tumor aggressiveness. In this study, we focused on the correlation between MMP9 expression and clear cell renal carcinoma (ccRCC). Through the Gene Expression Omnibus (GEO) database, the Cancer Genome Atlas (TCGA) database and immunohistochemical (IHC) staining, we observed that compared with adjacent normal renal tissues, in ccRCC tissues the mRNA and protein levels of MMP9 were enhanced, and the mRNA levels of GTP-binding protein smg p21B(RAP1B), B rapidly accelerated fibrosarcoma (RAF), methyl ethyl ketone2 (MEK2), extracellular regulated protein kinases1 (ERK1), ERK2, v-ets avian erythroblastosis virus E26 oncogene homolog1 (ETS1) and ETS2 also increased. The Kaplan–Meier survival analysis suggested that high MMP9 expression was an unfavorable prognostic biomarker for ccRCC patients. Our results indicated that the increased expression level of MMP9 in ccRCC may be due to the activation of the Mitogen-activated protein kinases (MAPK)/ERK signaling pathway, and MMP9 may be an attractive target for ccRCC therapy.
Background
Streptococcus agalactiae or group B Streptococcus (GBS) is a leading infectious cause of neonatal morbidity and mortality. It is essential to establish a robust method for the rapid and ultra-sensitive detection of GBS in pregnant women with premature rupture of membrane (PROM).
Methods
This study developed a CRISPR-GBS assay that combined the advantages of the recombinase polymerase amplification (RPA) and CRISPR/Cas12a system for GBS detection. The clinical performance of the CRISPR-GBS assay was assessed using vaginal or cervical swabs that were collected from 179 pregnant women with PROM, compared in parallel to culture-based matrix-assisted laser desorption ionization time-of-flight mass spectrometry (culture-MS) method and real-time quantitative polymerase chain reaction (qPCR) assay.
Results
The CRISPR-GBS assay can be completed within 35 min and the limit of detection was as low as 5 copies μL−1. Compared with the culture-MS, the CRISPR-GBS assay demonstrated a sensitivity of 96.64% (144/149, 95% confidence interval [CI] 92.39–98.56%) and a specificity of 100% (30/30, 95% CI 88.65–100%). It also had a high concordance rate of 98.88% with the qPCR assay.
Conclusions
The established CRISPR-GBS platform can detect GBS in a rapid, accurate, easy-to-operate, and cost-efficient manner. It offered a promising tool for the intrapartum screening of GBS colonization.
Background
Streptococcus agalactiae or group B Streptococcus (GBS) is a leading infectious cause of neonatal morbidity and mortality. This study aims to establish a novel method, termed as the CRISPR-GBS assay, for the rapid and sensitive detection of GBS that is based on the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas) system.
Results
The CRISPR-GBS assay detected GBS in the samples within 35 min. The limit of detection was as low as 5 copies/µL and showed no cross-reactivity with other microorganisms. The clinical performance was assessed using vaginal or cervical swab samples that were collected from 179 pregnant women with premature rupture of membrane. Compared with the culture-based matrix-assisted laser desorption ionization time-of-flight mass spectrometry method, the CRISPR-GBS assay demonstrated a sensitivity of 96.64% (144/149, 95% confidence interval [CI] = 92.39–98.56%) and a specificity of 100% (30/30, 95% CI = 88.65–100%). It also had a high concordance rate of 98.88% with the real-time fluorescence polymerase chain reaction assay.
Conclusions
The CRISPR-GBS assay can be used for rapid and high-sensitivity detection of GBS in a simple and cost-efficient manner; thus, it offers a novel method for intrapartum screening.
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