Clinical evaluation of rapid, simple and inexpensive electrochemical assay using LAMP amplification for detection of two most oncogenic human papillomavirus types, HPV16 and HPV18.
Cancer is a genetic disease induced by mutations in DNA, in particular point mutations in important driver genes that lead to protein malfunctioning and ultimately to tumorigenesis. Screening for the most common DNA point mutations, especially in such genes as TP53, BRCA1 and BRCA2, EGFR, KRAS, or BRAF, is crucial to determine predisposition risk for cancer or to predict response to therapy. In this review, we briefly depict how these genes are involved in cancer, followed by a description of the most common techniques routinely applied for their analysis, including high-throughput next-generation sequencing technology and less expensive low-throughput options, such as real-time PCR, restriction fragment length polymorphism, or high resolution melting analysis. We then introduce benefits of electrochemical biosensors as interesting alternatives to the standard methods in terms of cost, speed, and simplicity. We describe most common strategies involved in electrochemical biosensing of point mutations, relying mostly on PCR or isothermal amplification techniques, and critically discuss major challenges and obstacles that, until now, prevented their more widespread application in clinical settings.
Regionální centrum aplikované molekulární onkologie, Masarykův onkologický ústav, Brno Souhrn Východiska: Dlouhé nekódující RNA (long non-coding RNA-lncRNA) jsou RNA molekuly o délce větší než 200 nukleotidů, které ovlivňují řadu fyziologických funkcí a mají významnou regulační roli v buňkách. Jejich hladiny jsou často změněny u různých malignit, a představují tak slibný bio marker pro dia gnostiku, prognózu nebo rekurenci nádorových onemocnění. Díky důležitosti těchto molekul výrazně roste i počet publikací na toto téma. Mezi nejčastěji studované lncRNA patří např. HOTAIR, MALAT1 a PCA3. Cíl: V současné době jsou vyvíjeny různé metody pro analýzu či detekci lncRNA, obvykle založené na optických metodách pro detekci mediátorové RNA (mRNA), např. polymerázová řetězová reakce s reverzní transkripcí, fluorescenční in situ hybridizace nebo sekvenování nové generace. Je však potřeba dbát na rozdíly ve struktuře mRNA a lncRNA. V této práci popisujeme nejenom standardní metody, ale i nové přístupy pro detekci lncRNA zahrnující např. chemiluminescenční a elektrochemické techniky. Závěr: I navzdory pokrokům a velkému množství publikovaných prací existuje pouze jeden schválený dia gnostický test založený na detekci lncRNA, a to PCA3 test pro diagnostiku karcinomu prostaty analýzou moči. Ostatní jsou v současnosti pouze ve fázi vývoje a bude potřeba je validovat. Dia gnostika založená na lncRNA i tak skýtá obrovský potenciál, a je proto velmi pravděpodobné, že se v blízké době objeví další dia gnostické testy cílící na jiné typy lncRNA. Klíčová slova dlouhá nekódující RNA-nádorové bio markery-karcinogeneze-bio senzory Summary Background: Long non-coding RNAs (lncRNA) are more than 200-nucleotide-long RNA molecules that affect multiple physiologic phenomena and have important regulatory functions in cells. Their levels are often altered in various malignancies, thus they represent a potential biomarker for the diagnostics, prognosis or recurrence of cancer. Their importance has recently led to an enormous increase in a number of publications on the subject. The most frequently studied lncRNAs are HOTAIR, MALAT1 and PCA3. Aim: Numerous methods are currently being developed for the analysis or detection of lncRNA. They are mostly based on optical methods used for the detection of messenger RNAs, including polymerase chain reaction with reverse transcription, fluorescence in situ hybridisation or next-generation sequencing, but caution must be taken due to their structural differences. Here, we describe not only standard but also novel techniques for lncRNA detection, including chemiluminescent and electrochemical techniques. Conclusion: Despite the great progress and plethora of papers on this topic, there is only one single approved lncRNA-based diagnostic test, a PCA3 test for the diagnosis of prostate cancer from the patient's urine. All other tests are only in their research phase and need to be validated. Nevertheless, lncRNA diagnostics offer enormous potential and thus it is highly probable that other diagnostic tests o...
PUB were compared to the postsurgical pathological specimen or follow-up for non-operated tumors.
ResultsFrom November 2016 to February 2022, 34 patients were included. Based on surgical specimen (n=23) or follow up (n=11, including 4 metastasis), the final diagnoses were: 11 sarcomas and 23 non-sarcomas including 22 LM and one inflammatory myofibroblastic tumor [JS1] . The median followup was 12 months (IQR: 6-37). The diagnostic accuracy of M-PUB and MCGH-PUB were 94% and 100%. The sensitivity, specificity and Negative Predictive Value of MCGH-PUB were 100%, 100% and 100%. A high GI was significantly associated with malignancy (p<0•001). Genomic analyses allowed correct malignancy upgrade for four tumors after suspicious microscopic examination. There was no PUB complication and no dissemination on the biopsy track. Conclusion MCGH-PUB is safe and accurate to discriminate pre operatively benign tumors from uterine sarcoma.
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