Compositions and methods for detection of hepatitis A virus nucleic acid
View Patent ↗Nucleic acid oligomeric sequences and in vitro nucleic acid amplification and detection methods for detecting the presence of HAV RNA sequences in samples are disclosed. Kits comprising nucleic acid oligomers for amplifying and detecting HAV nucleic acid sequences are disclosed.
1. A combination of at least two oligomers for amplifying a hepatitis A virus (HAV) target region, wherein at least one amplification oligomer is selected from: an oligomer of about 20 to 30 contiguous nucleotides contained in the sequence of SEQ ID NO: 151 and which contains at least the sequence of SEQ ID NO:152; and wherein an additional amplification oligomer is selected from an oligomer of about 21-29 contiguous nucleotides contained in SEQ ID NO:165 and which contains at least the sequence of SEQ ID NO:166.
2. The combination of at least two oligomers of claim 1 , selected from the group consisting of: SEQ ID NO:75; SEQ ID NO:76; SEQ ID NO:77; SEQ ID NO:78; SEQ ID NO:104; SEQ ID NO:105; SEQ ID NO:106; SEQ ID NO:107; SEQ ID NO:108; SEQ ID NO:152; SEQ ID NO:153; SEQ ID NO:165; SEQ ID NO:166; SEQ ID NO:167; and SEQ ID NO:168.
3. The combination of at least two oligomers of claim 1 , further comprising at least one capture probe oligomer selected from the group consisting of SEQ ID NOS: 8, 9, 10, 11, 12, 13 and 14 and optionally linked to a moiety that allows it to bind a solid support.
4. The combination of at least two oligomers of claim 1 , further comprising at least one detection probe oligomer selected from the group consisting of SEQ ID NOS: 129, 130 and 131 and SEQ ID NO:136.
5. The combination of at least two oligomers according to claim 1 , wherein said combination is packaged in a kit.
6. The combination of at least two oligomers of claim 1 , wherein the oligomer contained within SEQ ID NO:151 and containing at least the sequence of SEQ ID NO:152, further includes a five prime promoter.
7. The combination of at least two oligomers of claim 6 , wherein the oligomer contained within SEQ ID NO:151 and containing at least the sequence of SEQ ID NO:152 and further including a five prime promoter is selected from the group consisting of SEQ ID NOS:45, 46, 47 and 48.
8. The combination of at least two oligomers of claim 1 , wherein the oligomer that is contained within SEQ ID NO:165 and includes at least the sequence of SEQ ID NO:166, further comprises a five primer promoter sequence.
9. The combination of at least two oligomers of claim 1 , comprising SEQ ID NO:46 or SEQ ID NO:76 and further comprising SEQ ID NO:108.
10. The combination of at least two oligomers of claim 1 , comprising SEQ ID NO:46 or SEQ ID NO:76 and further comprising SEQ ID NO:107.
11. The combination of at least two oligomers of claim 1 , further comprising one or more of a capture oligomer, an amplification oligomer, and a detection oligomer targeting a Parvovirus nucleic acid.
12. The combination of at least two oligomers of claim 1 , wherein two of the at least two oligomers are selected from the group consisting of;
(a) a first oligomer contained within SEQ ID NO:165 and including at least the sequence of SEQ ID NO:166 and a second oligomer that is SEQ ID NO:46 or SEQ ID NO:76;
(b) a first oligomer contained within SEQ ID NO:165 and including at least the sequence of SEQ ID NO:166 and a second oligomer that is SEQ ID NO:48 or SEQ ID NO:78;
(c) a first oligomer contained within SEQ ID NO:151 and containing at least the sequence of SEQ ID NO:152, and optionally includes a five prime promoter, and a second oligomer that is SEQ ID NO:107; and
(d) a first oligomer contained within SEQ ID NO:151 and containing at least the sequence of SEQ ID NO:152 , and optionally includes a five prime promoter, and a second oligomer that is SEQ ID NO:108.
13. A method of detecting the presence of hepatitis A virus (HAV) in a sample comprising the steps of:
purifying HAV nucleic acid from other components in a sample suspected of containing HAV;
amplifying an HAV target sequence in the purified HAV nucleic acid, or a HAV cDNA made therefrom, by using an in vitro amplification reaction that includes at least two amplification oligomers, wherein at least one amplification oligomer is selected from an oligomer of about 20 to 30 contiguous nucleotides contained in the sequence of SEQ ID NO:151 and which contains at least the sequence of SEQ ID NO:152; and wherein an additional amplification oligomer is selected from-an oligomer of about 21-29 contiguous nucleotides contained in SEQ ID NO:165 and which contains at least the sequence of SEQ ID NO:166; and
performing a detection reaction to identify the presence or absence of an amplified HAV product as an indication of the presence or absence of the HAV nucleic acid in the sample.
14. The method of claim 13 , wherein the purifying step contacts the sample with at least one capture probe oligomer selected from the group consisting of SEQ ID NOS: 8, 9, 10, 11, 12, 13 and 14 linked to a moiety that allows it to bind a solid support.
15. The method of claim 13 , wherein the amplifying step uses at least two oligomers selected from the group consisting of: SEQ ID NO:75; SEQ ID NO:76; SEQ ID NO:77; SEQ ID NO:78; SEQ ID NO:104; SEQ ID NO:105; SEQ ID NO:106; SEQ ID NO:107; SEQ ID NO:108; SEQ ID NO:152; SEQ ID NO:153; SEQ ID NO:165; SEQ ID NO:166; SEQ ID NO:167; and SEQ ID NO:168.
16. The method of claim 13 , wherein said nucleic acid detection oligomer comprises a sequence selected from the group consisting of SEQ ID NO:129, SEQ ID NO:130 and SEQ ID NO:136.
17. The method of claim 13 , wherein the amplifying step uses an oligomer contained within SEQ ID NO:151 and containing at least the sequence of SEQ ID NO:152, further comprises a five prime promoter.
18. The method of claim 17 , wherein the oligomer is selected from the group consisting of SEQ ID NOS:45, 46, 47 and 48.
19. The method of claim 13 , wherein the oligomer contained within SEQ ID NO:165 and that includes at least the sequence of SEQ ID NO:166, further comprises a five prime promoter sequence.
20. The method of claim 13 , wherein the amplification oligomer contained in the sequence of SEQ ID NO:151 and containing the sequence of SEQ ID NO:152 is SEQ ID NO:46 or SEQ ID NO:76 and wherein the amplification oligomer contained in SEQ ID NO:165 and containing SEQ ID NO:166 is SEQ ID NO:108.
21. The method of claim 13 , wherein the amplification oligomer contained in the sequence of SEQ ID NO:151 and containing the sequence of SEQ ID NO:152 is SEQ ID NO:46 or SEQ ID NO:76 and wherein the amplification oligomer contained in SEQ ID NO:165 and containing SEQ ID NO:166 is SEQ ID NO:107.
22. The method of claim 13 , further comprising one or more of a capture oligomer, an amplification oligomer, and a detection oligomer targeting a Parvovirus nucleic acid.
23. The method of claim 13 , wherein the sample is a biological sample collected from humans.
24. The method of claim 13 , wherein the at least two oligomers used at the amplifying step includes combinations of two amplification oligomers selected from the group consisting of:
(a) a first oligomer contained within SEQ ID NO:165 and including at least the sequence of SEQ ID NO:166 and a second oligomer that is SEQ ID NO:46 or SEQ ID NO:76;
(b) a first oligomer contained within SEQ ID NO:165 and including at least the sequence of SEQ ID NO:166 and a second oligomer that is SEQ ID NO:48 or SEQ ID NO:78;
(c) a first oligomer contained within SEQ ID NO: 151 and containing at least the sequence of SEQ ID NO:152, and optionally includes a five prime promoter, and a second oligomer that is SEQ ID NO:107; and
(d) a first oligomer contained within SEQ ID NO:151 and containing at least the sequence of SEQ ID NO:152, and optionally includes a five prime promoter, and a second oligomer that is SEQ ID NO:108.
25. The method of claim 23 , wherein the sample collected from a human is a plasma sample.
26. The method of claim 23 , wherein the sample collected from a human is a sample used for the preparation of therapeutic factors isolated from humans.