The most recent diagnostic methods employed in determining an MTHFR deficiency starts with a basic test for the concentration of homocysteine in plasma using either whole blood samples or according to Fowler, B. and Jakobs, C., 1998, complete homocysteine measurement in the cell-free amniotic fluid can also work. This can be quantified with the help of a traditional amino acid chromatography or as is more regular as of late, through the use of a radioenzymatic procedure that measures the sum of homocysteine levels, such as protein-bound, free, oxidized and reduced structures (Tonetti et al 2000).
When the homocysteine levels are discovered to be higher (albeit ordinarily, it is supposed to be completely absent), an immediate fibroblasts and/or blood platelets analysis is conducted to measure the precise MTHFR activity level as already portrayed by Kutzbach and Stokstad (1971). In the event that the activity level of the MTHFR is observed to be unusually low, then an initial isolation of the DNA from cultured fibroblasts is done for use in the mutation evaluation. The precise mutations can then be decided for specific exons as proposed by Goyette et al (1998) or by having the purified PCR fragments directly sequenced and carried out by a programmed sequencer utilizing color marked terminators. According to Tonetti et al., 2000, it is possible to verify the mutations discovered through direct sequencing of PCR fragments of diverse exons by making use of restriction enzymes, for example, Hinf I for exon 4, Hha I for exon 6, and so forth.
Direct sequencing is also great at identifying the common MTHFR alleles. Nonetheless, since these alleles generate (C677T) or eradicate (A1298C) particular restriction sites, it is easier to identify them first by digesting the polymerase chain reaction-amplified genomic DNA using the suitable restriction enzyme (Hinf I for the C677T allele and Mbo II for the A1298C allele) and after that having the fragments separated by gel electrophoresis. According to Botto and Yang, 2000, the given process has been carried out on DNA gotten from diverse sources, such as amniotic fluid, blood, and congealed blood spots.
It ought to be known, nonetheless, that diagnosis of chronic MTHFR deficiency on the premise of just DNA tests is inadequate. This is on the grounds that there is not one mutation connected to this chronic strain of the ailment, and as such checking for identifiable mutations may prevent the recognition of an unidentifiable chronic mutation.
At the point when this deficiency is diagnosed, MTHFR treatment can commence. For mild deficiency, MTHFR treatment often involves just folate supplementation and eating regimen control. This is because earlier researches show that folates will shield mild MTHFR mutant proteins from FAD loss, and as such reestablishing enzyme capabilities to almost standard levels (Guenther and Sheppard et al, 1999).
An aggressive approach needs to be taken towards the treatment of severe deficiency of MTHFR, albeit and, moreover, such treatment procedure is not generally compelling. Severe deficiency of MTHFR needs treatments that utilize a blend of therapies with the sole aim of lowering homocysteine levels and increasing any lingering enzyme activity. The principal form of treatment involves making use of folate compounds, such as folinic or folic acid (Rosenblatt, 1995). Inside the human body, these compounds function by boosting enzyme activity, allegedly via their capacity to shield the protein catalyst from losing its essential FAD cofactor (Guenther and Sheppard, et. al, 1999). Another therapy used in this deficiency is methyltetrahydrofolate treatment, which is tasked with the function of making up for the inability of methylenetetrahydrofolate reductase to generate this product. Further utilized as a part of the treatment procedure are two compounds tasked with making up for the low methionine levels that cause MTHFR deficiency: 1) methionine itself, and 2) betaine, a substrate for betaine methyltransferase, a compound that gives another option for the conversion of homocysteine to methionine (Rosenblatt, 1995).