Hepatitis A virus nucleotide sequences, recombinant proteins and uses thereof
View Patent ↗Hepatitis A virus primers and probes derived from the capsid proteins and junction between the capsid precursor P1 and 2A of the HAV genome are disclosed. Also disclosed are nucleic acid-based assays using the primers and probes, antigen detection of HAV, and immunoassay for detecting the antibodies that bind to HAV.
1. An isolated polynucleotide comprising:
(a) a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 15;
(b) a sequence complementary to the sequence of (a); or
(c) a fragment of either of the sequences in (a) or (b) wherein the fragment comprises at least 15 contiguous nucleotides of SEQ ID NO: 15 or of the complement of SEQ ID NO: 15.
2. The polynucleotide of claim 1 , wherein the nucleotide sequence comprises the sequence of SEQ ID NO: 15.
3. A method for detecting Hepatitis A virus (HAV) infection in a biological sample, the method comprising:
(a) isolating nucleic acid from a biological sample suspected of containing Hepatitis A virus (HAV) RNA, wherein said nucleic acid comprises a target sequence;
(b) reacting the HAV nucleic acid with a detectably labeled probe sufficiently complementary to and capable of hybridizing with the target sequence, wherein the probe is a polynucleotide according to claim 1 , and further wherein said reacting is done under conditions that provide for the formation of a probe/target sequence complex; and
(c) detecting the presence or absence of label as an indication of the presence or absence of the target sequence.
4. The method of claim 3 , wherein the probe comprises a detectable label at the 5′-end and/or at the 3′-end.
5. The method of claim 3 , wherein the detectable label is a fluorescent label selected from the group consisting of 6-carboxyfluorescein (6-FAM), tetramethyl rhodamine (TAMRA), and 2′, 4′, 5′, 7′,-tetrachloro-4-7-dichlorofluorescein (TET).
6. A method for detecting Hepatitis A virus (HAV) infection in a biological sample, the method comprising:
isolating nucleic acids from a biological sample suspected of containing HAV;
amplifying the nucleic acids using at least two primers wherein each of the primers is not more than about 50 nucleotides in length and comprises a nucleotide sequence of at least 10 contiguous nucleotides from a polynucleotide according to claim 1 or a nucleotide sequence having 90% sequence identity thereto, wherein each of the two primer is sufficiently complementary to a portion of the sense and antisense strands, respectively, of the isolated nucleic acid to hybridize therewith; and
detecting the of the amplified nucleic acids as an indication of the presence or absence of HAV in the sample.