Compositions and methods for internal controls of microvesicle isolations
AssigneeExosome Diagnostics, Inc.
InventorsGeorg STOLL, Daniel ENDERLE, Mikkel NOERHOLM, Johan Karl Olov SKOG
Life of this patent
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Abstract
The present disclosure provides compositions, methods and kits for internal controls of microvesicle isolations. The compositions, methods and kits can comprise enveloped viruses, including, but not limited to, inactive mouse hepatitis virus (MHV).
Claims41
A method comprising:
- (a) adding a known quantity of control particles comprising at least one control nucleic acid and/or at least one control protein to a biological sample, wherein the control particles comprise an enveloped virus;
- (b) isolating microvesicles and the control particles from the biological sample;
- (c) extracting nucleic acids and/or protein from the microvesicles and the control particles;
- (d) assaying the amount of the at least one control nucleic acid and/or the at least one control protein recovered, thereby determining the amount of control particles recovered;
- (e) determining that the amount of control particles calculated in step (d) is within a predetermined range of values;
- and
- (f) subjecting the nucleic acids and/or protein from step (c) to further analysis if the amount of control particle calculated in step (d) is within the predetermine range of values.
The method of any of the preceding claims, wherein the non-human animal enveloped virus is an inactivated non-human animal enveloped virus.
The method of any of the preceding claims, wherein the enveloped virus is a human enveloped virus.
The method of any of the preceding claims, wherein the human enveloped virus is inactivated human enveloped virus.
The method of any of the preceding claims, wherein the inactivated non-human animal enveloped virus is mouse hepatitis virus (MHV).
The method of any of the preceding claims, wherein the inactivated non-human animal enveloped virus is bovine viral diarrhea virus (BVDV).
The method of any of the preceding claims, wherein the inactivated non-human animal enveloped virus is transmissible gastroenteritis coronavirus (TGEV).
The method of any of the preceding claims, wherein the biological sample is a bodily fluid sample.
The method of any of the preceding claims, wherein the bodily fluid sample is urine, blood, cerebrospinal fluid or serum.
The method of any of the preceding claims, wherein the at least one control nucleic acid comprises RNA.
The method of any of the preceding claims, wherein the nucleic acids from step (c) comprises RNA.
The method of any of the preceding claims, wherein the nucleic acids from step (c) comprises DNA.
The method of any of the preceding claims, wherein the nucleic acids from step (c) comptises RNA and DNA.
The method of any of the preceding claims, wherein assaying the amount of the at least one control nucleic acid recovered comprises determining the expression level or copy number of the control nucleic acid.
The method of any of the preceding claims, wherein assaying the expression level or copy number of the control nucleic acid comprises reverse transcribing the control nucleic acid.
The method of any of the preceding claims, wherein assaying the expression level or copy number of the control nucleic acid comprises quantitative PCR (qPCR).
The method of any of the preceding claims, wherein step (f) comprises determining the presence, absence or level of at least one biomarker associated with a disease or medical condition for diagnosing, prognosing, or monitoring the disease or medical condition.
The method of any of the preceding claims, wherein determining the presence, absence or level of at least one biomarker associated with a disease or medical condition for diagnosing, prognosing, or monitoring the disease or medical condition comprises reverse transcription.
The method of any of the preceding claims, wherein determining the presence, absence or level of at least one biomarker associated with a disease or medical condition for diagnosing, prognosing, or monitoring the disease or medical condition comprises quantitative PCR.
A method comprising:
- (a) isolating microvesicles from a biological sample;
- (b) adding a known quantity of control particles comprising at least one control nucleic acid and/or at least one control protein to the isolated microvesicles, wherein the control particles comprise an enveloped virus;
- (c) extracting nucleic acids and/or protein from the isolated microvesicles and the control particles;
- (d) assaying the amount of the at least one control nucleic acid and/or the at least one control protein recovered, thereby determining the amount of control particles recovered;
- (e) determining that the amount of control particles calculated in step (d) is within a predetermined range of values;
- and
- (f) subjecting the nucleic acids and/or protein from step (c) to further analysis if the amount of control particle calculated in step (d) is within the predetermine range of values.
The method of any of the preceding claims, wherein the enveloped virus is a non-human animal enveloped virus.
The method of any of the preceding claims, wherein the non-human animal enveloped virus is an inactivated non-human animal enveloped virus.
The method of any of the preceding claims, wherein the enveloped virus is a human enveloped virus.
The method of any of the preceding claims, wherein the human enveloped virus is inactivated human enveloped virus.
The method of any of the preceding claims, wherein the inactivated non-human animal enveloped virus is mouse hepatitis virus (MHV).
The method of any of the preceding claims, wherein the inactivated non-human animal enveloped virus is Bovine viral diarrhea virus (BVDV).
The method of any of the preceding claims, wherein the inactivated non-human animal enveloped virus is transmissible gastroenteritis coronavirus (TGEV).
The method of any of the preceding claims, wherein the biological sample is a bodily fluid sample.
The method of any of the preceding claims, wherein the bodily fluid sample is urine, blood, cerebrospinal fluid or serum.
The method of any of the preceding claims, wherein the at least one control nucleic acid comprises RNA.
The method of any of the preceding claims, wherein the nucleic acids from step (c) comprises RNA.
The method of any of the preceding claims, wherein the nucleic acids from step (c) comprises DNA.
The method of any of the preceding claims, wherein the nucleic acids from step (c) comprises RNA and DNA.
The method of any of the preceding claims, wherein assaying the amount of the at least one control nucleic acid recovered comprises assaying the expression level or copy number of the control nucleic acid.
The method of any of the preceding claims, wherein assaying the expression level or copy number of the control nucleic acid comprises reverse transcribing the control nucleic acid.
The method of any of the preceding claims, wherein assaying the expression level or copy number of the control nucleic acid comprises quantitative PCR (qPCR).
The method of any of the preceding claims, wherein step (f) comprises determining the presence, absence or level of at least one biomarker associated with a disease or medical condition for diagnosing, prognosing, or monitoring the disease or medical condition.
The method of any of the preceding claims, wherein determining the presence, absence or level of at least one biomarker associated with a disease or medical condition for diagnosing, prognosing, or monitoring the disease or medical condition comprises reverse transcription.
The method of any of the preceding claims, wherein determining the presence, absence or level of at least one biomarker associated with a disease or medical condition for diagnosing, prognosing, or monitoring the disease or medical condition comprises quantitative PCR.
The method of any of the preceding claims, wherein the quantitative PCR is reverse transcription quantitative PCR (RT-qPCR).
Dependencies are inferred from the claim text.