Phytohemagglutinin stimulation of lymphocytes improves mutation analysis of carbamoylphosphate synthetase 1

Molecular Genetics and Metabolism
Rita KretzJohannes Häberle

Abstract

Carbamoylphosphate synthetase 1 (CPS1) is the first enzyme of the urea cycle. CPS1 deficiency is a rare autosomal-recessively inherited disorder that can lead to life-threatening hyperammonemia. Since there is no reliable biochemical marker for this disease, diagnosis relies on molecular means which is often done by RNA-based mutation analysis. Skin fibroblasts have been frequently used as a source of RNA while peripheral blood cells do not yield sufficient amounts of specific RNA. To avoid the costly and laborious use of cultured fibroblasts, we tried to use stimulated lymphocytes as an alternative. This was effectively achieved by short-term culture of full heparin blood in the presence of phytohemagglutinin. Hereby, subsequent reverse transcriptase-PCR of the CPS1 transcript became feasible and allowed to detect 16 different mutations (10 missense, 3 deletions, 2 nonsense, 1 duplication; 7 novel mutations) in 14 consecutive patients with CPS1 deficiency. When compared to retrospective data on cultured fibroblasts, the adapted method allowed substantial shortening of the median time to diagnosis (24 days versus 122 days, respectively). Besides disease causing mutations, we detected CPS1 transcript variants including one crypt...Continue Reading

References

Apr 1, 1970·Experimental Cell Research·J P Monjardino, A J MacGillivray
Jun 1, 1995·Nature Genetics·F B HogervorstH Meijers-Heijboer
Jul 20, 2001·Human Mutation·A JakubowskaJ Lubinski
Dec 25, 1964·Science·J H ROBBINS

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Citations

Feb 26, 2016·Genetics in Medicine : Official Journal of the American College of Medical Genetics·Carmen Diez-FernandezJohannes Häberle
Mar 11, 2017·Expert Opinion on Therapeutic Targets·Carmen Diez-Fernandez, Johannes Häberle
Apr 27, 2017·Journal of Clinical Laboratory Analysis·Guoqing ZhangJun Bu
Apr 16, 2019·Journal of Inherited Metabolic Disease·Johannes HäberleCarlo Dionisi-Vici
Oct 5, 2018·Orphanet Journal of Rare Diseases·Sunita Bijarnia-MahayIshwar C Verma

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