Disclosed herein are in vitro methods of multiple staining and slide preparation for a cytopathological sample, such as a urine sample. These methods enable simultaneous staining of biomarkers and cytopathological stains to greatly enhance confidence in identifying atypical cells such as cancer cells. Also disclosed herein are methods of using said staining and preparation methods for the detection of bladder cancer using urine samples from a patient. These methods offer increased sensitivity and specificity over conventional bladder cancer detection methods. Also disclosed herein are the urinary exfoliated cells stained according to the methods provided herein.

A nucleic acid (NA) detection method combines ultra-specific probe, on-chip isotachophoresis (ITP) which can separate single strand and double strand NAs, and enzyme amplification. The ITP device has a sieving matrix between the LE (leading electrolyte) and TE (trailing electrolyte) reservoirs, for separating double-strand and single-strand NAs. The LE or TE reservoir also contains a spacer ion having a mobility between the LE and the TE. The sample and a double-strand NA probe is added to the TE reservoir, the probe being formed of a protector strand modified with a fluorescent molecule and a complement strand, where the protector strand is released in the presence of the target NA. Fluorescent signal is detected downstream of the sieving matrix. Alternatively, the protector strand is modified with an enzyme and a single-strand NA modified with a substrate of the enzyme is added to the reaction mixture downstream of the sieving matrix.

A nucleic acid (NA) detection method combines ultra-specific probe, on-chip isotachophoresis (ITP) which can separate single strand and double strand NAs, and enzyme amplification. The ITP device has a sieving matrix between the LE (leading electrolyte) and TE (trailing electrolyte) reservoirs, for separating double-strand and single-strand NAs. The LE or TE reservoir also contains a spacer ion having a mobility between the LE and the TE. The sample and a double-strand NA probe is added to the TE reservoir, the probe being formed of a protector strand modified with a fluorescent molecule and a complement strand, where the protector strand is released in the presence of the target NA. Fluorescent signal is detected downstream of the sieving matrix. Alternatively, the protector strand is modified with an enzyme and a single-strand NA modified with a substrate of the enzyme is added to the reaction mixture downstream of the sieving matrix.

Under one aspect, a composition includes a substrate; a first polynucleotide coupled to the substrate; a second polynucleotide hybridized to the first polynucleotide; and a catalyst coupled to a first nucleotide of the second polynucleotide, the catalyst being operable to cause a chemiluminogenic molecule to emit a photon. Under another aspect, a method includes providing a catalyst operable to cause a first chemiluminogenic molecule to emit a photon; providing a substrate; providing a first polynucleotide coupled to the substrate; hybridizing a second polynucleotide to the first polynucleotide; coupling a first quencher to a first nucleotide of the second polynucleotide; and inhibiting, by the first quencher, photon emission by the first chemiluminogenic molecule.

Provided are methods of isolating RNA from a biological sample, methods and means for determining the presence of particular RNA splice-form variants in a biological sample, methods and means for determining the relative ratio of RNA ratios in a biological sample, and methods and means for predicting the progression of precancerous cervical lesions.

Provided are methods of isolating RNA from a biological sample, methods and means for determining the presence of particular RNA splice-form variants in a biological sample, methods and means for determining the relative ratio of RNA ratios in a biological sample, and methods and means for predicting the progression of precancerous cervical lesions.

This document provides methods and materials for detecting target nucleic acid. For example, methods and materials for detecting the presence or absence of target nucleic acid, methods and materials for detecting the amount of target nucleic acid present within a sample, kits for detecting the presence or absence of target nucleic acid, kits for detecting the amount of target nucleic acid present within a sample, and methods for making such kits are provided.

This document provides methods and materials for detecting target nucleic acid. For example, methods and materials for detecting the presence or absence of target nucleic acid, methods and materials for detecting the amount of target nucleic acid present within a sample, kits for detecting the presence or absence of target nucleic acid, kits for detecting the amount of target nucleic acid present within a sample, and methods for making such kits are provided.

This document provides methods and materials for assessing RNA expression. For example, methods and materials for detecting the presence, absence, or amount of target nucleic acid (e.g., target RNA or target cDNA produced from target RNA), kits for detecting the presence, absence, or amount of target nucleic acid (e.g., target RNA or target cDNA produced from target RNA), and methods for making such kits are provided.

This document provides methods and materials for assessing RNA expression. For example, methods and materials for detecting the presence, absence, or amount of target nucleic acid (e.g., target RNA or target cDNA produced from target RNA), kits for detecting the presence, absence, or amount of target nucleic acid (e.g., target RNA or target cDNA produced from target RNA), and methods for making such kits are provided.