Compositions and methods for detecting allogeneic matter in a subject
The present disclosure provides a panel of nucleic acid molecule primers specific for HLA-specific alleles and other genetic polymorphisms, which are useful for quantitatively amplifying these markers to detect, diagnose, and monitor individuals who have or are at risk of certain disease conditions, such as autoimmune disease, proliferative disease, infectious disease, allograft rejection, or pregnancy-related pathologies.
1. A kit for use in carrying out a method for detecting microchimerism, the kit comprising one or more specific nucleic acid molecule compositions comprising a forward nucleic acid molecule comprising the sequence set forth in SEQ ID NO: 64, a reverse nucleic acid molecule comprising the sequence set forth in SEQ ID NO: 65, and a probe nucleic acid molecule comprising the sequence set forth in SEQ ID NO: 66;
wherein the method comprises:
(a) amplifying nucleic acid molecules of a target biological sample using the specific nucleic acid molecule compositions;
(b) amplifying nucleic acid molecules of a control biological sample using the specific nucleic acid molecule compositions of step (a);
(c) comparing the amount of amplified nucleic acid molecules with a reference to quantitate specific nucleic acid molecule markers in the samples;
wherein each specific nucleic acid molecule composition amplifies a SE-LR genetic polymorphism; and
wherein the presence of certain specific amplified markers in the target biological sample indicates the presence of microchimerism.
2. A kit for use in detecting incipient allograft rejection from a biological sample of a subject, the kit comprising one or more specific nucleic acid molecule compositions comprising a forward nucleic acid molecule comprising the sequence set forth in SEQ ID NO: 64, a reverse nucleic acid molecule comprising the sequence set forth in SEQ ID NO: 65, and a probe nucleic acid molecule comprising the sequence set forth in SEQ ID NO: 66.
3. The kit of claim 1 , wherein the kit has a detection sensitivity of one chimeric genome in 10 5 host genomes.
4. The kit of claim 1 , wherein the biological sample is blood or serum.
5. The kit of claim 1 , wherein the amplifying comprises performing Q-PCR.
6. The kit of claim 2 , wherein the biological sample is blood or serum.
7. The kit of claim 2 , wherein the subject is a human.
8. The kit of claim 2 , wherein the subject has a known genotype.