Method for the specific detection and identification of retroviral nucleic acids/retroviruses in a specimen

A method for specific detection and identification of a retrovirus or retroviral nucleic acid in a specimen comprising the steps of:

• isolating at least one, DNA or RNA from the specimen;
• producing respective amplificates of the DNA or RNA by subjecting said at least one isolated DNA to PCR or isolated RNA to RT-PCR by using a primer mixture of forward and reverse primers, wherein the primer mixture comprises at least one of MOP-ABD or MOP-C and wherein MOP-ABD consists of a forward primer with a head at its 5′ end and a nucleotide sequence in accordance with SEQ. ID NO. 1 with 3456 degenerations, and a reverse primer with a head at its 5′ end and a nucleotide sequence in accordance with SEQ. ID NO.2 with 27648 degenerations;
• and wherein MOP-C, consists of a forward primer which is a nucleotide sequence with a head at its 5′ end and a nucleotide sequence in accordance with SEQ. ID NO. 3 with 3072 degenerations, and a reverse primer with a head at its 5′ end and a nucleotide sequence in accordance with SEQ. ID NO. 4 with 8192 degenerations;
• and wherein each of the heads defines a nucleotide sequence comprising an interface for a restriction enzyme and a clamp sequence at a 5′ end of the interface and which has a length not exceeding one half of a length of a complete nucleotide sequence of one of the forward or the reverse primer;
• purging the amplificates;
• detecting and identifying the presence of a retroviral nucleotide sequence of a retrovirus-specific reverse-transcriptase gene or a section thereof by subjecting the amplificates to reverse dot blot hybridization (RDBH) using immobilized RDBH probes, wherein each said probes includes at least one of synthetic oligonucleotide sequences corresponding to the retroviral nucleotide sequence of the retrovirus specific reverse transcriptase gene or a section thereof, and which do not overlap with nucleotide sequences of the forward and reverse primer of the primer mixture.

A diagnostic kit for the specific detection and identification of retroviral nucleic acids and/or retroviruses in an arbitrary specimen comprising:

• at least one of MOP-ABD or MOP-C and wherein MOP-ABD consists of a forward primer with a head at its 5′ end and a nucleotide sequence in accordance with SEQ. ID NO. 1 with 3456 degenerations, and a reverse primer with a head at its 5′ end and a nucleotide sequence in accordance with SEQ. ID NO.2 with 27648 degenerations;
• and wherein MOP-C, consists of a forward primer which is a nucleotide sequence with a head at its 5′ end and a nucleotide sequence in accordance with SEQ. ID NO. 3 with 3072 degenerations, and a reverse primer with a head at its 5′ end and a nucleotide sequence in accordance with SEQ. ID NO. 4 with 8192 degenerations;
• and wherein each of the heads defines a nucleotide sequence comprising an interface for a restriction enzyme and a clamp sequence at a 5′ end of the interface and which has a length not exceeding one half of a length of a complete nucleotide sequence of one of the forward or the reverse primer;
• and
• at least one reverse dot blot hybridization probe.