Molecular diagnosis of factor XIII deficiency, data from comprehensive coagulation laboratory in Iran

Abstract

: Diagnosis of factor XIII (FXIII) deficiency (FXIIID) as a rare bleeding disorder is a challenge worldwide. Thus, in the present study, we used different methods including two molecular methods for detection of FXIIID. This study was conducted on individuals suspected to FXIIID. All individuals were checked by two routinely used methods of clot solubility test in Iran and two other clot solubility tests as well as FXIII activity and antigen assays. Molecular analysis was performed by PCR-restriction fragment length polymorphism (PCR-RFLP) and tetra-primer amplification refractory mutation system (T-ARMS)-PCR for only FXIIID mutation in southeast Iran (p.Trp187Arg), previously associated with severe FXIIID. Out of 151 individuals, 26 had abnormal clot solubility test with all four methods. PCR-RFLP revealed that 27 patients were homozygotes for p.Trp187Arg, whereas 12 were heterozygotes. Molecular analysis revealed that in routinely used clot solubility combinations, two homozygotes (∼8%) were missed, whereas in two other combinations, one patient (∼4%) was missed. One false positive result was observed in routinely used methods, whereas further combinations don't have false positive. T-ARMS-PCR had three discrepancies with PCR-RFLP and sequencing confirmed that the results of T-ARMS-PCR were false. FXIII antigen assay diagnosed all homozygotes, whereas in FXIII activity assay, two homozygotes had higher than 5% FXIII activity that inconsistent with severe deficiency. It seems that clot solubility test is not enough sensitive and specific and molecular analysis is the most reliable method for detection of FXIIID in areas such Iran with one or few specific mutations.

A Unique Factor XIII Mutation in Southeastern Iran with an Unexpectedly High Prevalence: Khash Factor XIII

“…[10][11][12][13][14][15] Although the diagnosis of FXIII deficiency in Iran is still performed mostly by means of clot solubility tests, using 1% monochloroacetic acid (MCA) and 5M urea, several newer and improved diagnostic techniques, including FXIII activity assays and molecular analysis are growing in use in recent years. It has been shown that clot-solubility tests cannot detect all patients with severe FXIII deficiency in southeastern Iran, 16,17 which is similar to experience seen elsewhere. 18 Although UCB is the leading clinical symptom leading to a diagnosis of FXIII deficiency, both in Iran and worldwide, ICH is the main cause of death in these patients.…”

“…We have found that neither the clot solubility test nor the FXIII activity assay detects all patients with severe FXIII deficiency, while a molecular study with a simple polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) can instead detect all patients. 16 Comparatively, the tetra-primer amplification refractory mutation system-PCR (T-ARMS-PCR) as a cost-benefit molecular method has failed to detect all cases with FXIII deficiency. 16 All patients with confirmed FXIII deficiency prophylactically receive Fibrogammin P in a dose of 10 IU/ kg every 4 weeks from the time of diagnosis.…”

Expression and CpG island methylation pattern of MMP-2 and MMP-9 genes in patients with congenital factor XIII deficiency and intracranial hemorrhage

“…ICH is the main cause of morbidity and mortality in congenital FXIII deficiency. Due to the rarity of congenital FXIII deficiency, there is a paucity of data in the literature about the pathogenesis and underlying factors of ICH in this disorder [3][4][5][6]. According to the previous studies, MMP-2 and MMP-9 with gelatinase activity, have been broadly associated with BBB breakdown and ICH [11,21].…”

“…Congenital FXIII deficiency is a rare bleeding disorder (RBD) with an autosomal recessive pattern of inheritance and estimated incidence of 1 per 2 million in the general population. The disorder is more common in Iran, especially Khash city (Khash factor XIII) with an incidence of 1 homozygote per ∼500 individuals and 3.5% heterozygotes, in this area [3][4][5][6][7]. Based on residual plasma FXIII level in patients with FXIII deficiency, bleeding symptoms can range from mild to severe such as intracranial hemorrhage (ICH).…”

“…Furthermore, ICH is more common in congenital FXIII deficiency than any other congenital bleeding disorder. Occurrence of ICH in congenital FXIII deficiency can be affected by several factors, including residual plasma factor level, age of patient and prophylaxis regimen [3][4][5][6][7][8].…”