Autism spectrum disorder (ASD) is a highly heterogeneous neurodevelopmental disorder with many contributing risk genes and loci. To date, several intellectual disability (ID) susceptibility genes have frequently been identified in ASD. Here, whole exome sequencing was carried out on a proband with ASD and identified compound heterozygous mutations of the TRAPPC9, which plays a role in the neuronal NF-κB signaling pathway. These mutations consisted of a novel frameshift mutation (c.2415_2416insC, p.His806Profs^∗9) and a rare splice site mutation (c.3349+1G>A) that were segregated from an unaffected father and unaffected mother, respectively. These two heterozygous mutations were also identified in the patient’s older brother with ID. Quantitative RT-PCR revealed a significant reduction of TRAPPC9 transcript in two siblings. This study first describes compound heterozygous mutations of the TRAPPC9 gene in two siblings with ASD and ID, which is notable as only homozygous mutations or compound heterozygous for copy number variations and rare variant in this gene have been reported to date and associated with autosomal recessive intellectual disability. The two siblings carrying compound heterozygous TRAPPC9 mutations presented with ID, developmental delay, microcephaly and brain abnormalities similarly to the clinical features found in almost cases with homozygous TRAPPC9 mutation in previous studies. Together this study provides evidence that clinical manifestations of TRAPPC9 mutations as seen in our patients with ID and autism may be broader than previous case reports have indicated.

Chromosomal microarray (CMA) is now recognized as the first-tier genetic test for detection of copy number variations (CNVs) in patients with autism spectrum disorder (ASD). The aims of this study were to identify known and novel ASD associated-CNVs and to evaluate the diagnostic yield of CMA in Thai patients with ASD. The Infinium CytoSNP-850K BeadChip was used to detect CNVs in 114 Thai patients comprised of 68 retrospective ASD patients (group 1) with the use of CMA as a second line test and 46 prospective ASD and developmental delay patients (group 2) with the use of CMA as the first-tier test. We identified 7 (6.1%) pathogenic CNVs and 22 (19.3%) variants of uncertain clinical significance (VOUS). A total of 29 patients with pathogenic CNVs and VOUS were found in 22% (15/68) and 30.4% (14/46) of the patients in groups 1 and 2, respectively. The difference in detected CNV frequencies between the 2 groups was not statistically significant (Chi square = 1.02, df = 1, P = 0.31). In addition, we propose one novel ASD candidate gene, SERINC2, which warrants further investigation. Our findings provide supportive evidence that CMA studies using population-specific reference databases in underrepresented populations are useful for identification of novel candidate genes.

Microscopic examination is essential in urine analysis. This is a simple way to collect informative data but it is also labor-intensive, time-consuming, and requires experienced staff for accurate results and interpretation. Several automated urine analyzers have been introduced for urine analysis in medical laboratories. The aim of this study was to assess and compare the performance of the most common three automated urine analyzers, Cobas 6500, UN3000-111b and iRICELL 3000. and Methods: A total of 100 routine urine samples were used in the study. Results from the three machines were compared with the routine procedure results including physical, chemical and sediment analysis. There was good correlation of urine physical and chemical analyses between the three analyzers with an overall concordance level of more than 80%. For sediment analysis, the degree of concordance between manual analysis and the three instruments was very good to good for white blood cells, red blood cells and epithelial cells, and moderate for bacteria. There were fair to good agreements between manual microscopy and the three instruments, Cobas 6500, UN3000-111b and iRICELL 3000, for casts (Cohen’s kappa 0.42, 0.38 and 0.62, respectively). The three automated urine analyzers showed similar performances and good correlation with manual microscopy. The results of this study indicate that automated urine analyzers could be used for initial urine testing to reduce high workloads and to save time, but manual microscopic analysis by experienced staff is still necessary to classify urine sediments for confirmation, especially in pathologic specimens.

To identify the underlying genetic cause of autism spectrum disorder (ASD), we performed whole-exome sequencing in 10 unrelated Thai patients with ASD. We identified a novel heterozygous missense variant (c.425C>G, p.Pro142Arg) in the Engrailed 2 (EN2) gene in two patients. The G variant has never been reported in public databases and was absent in 100 Thai patients with ASD and 435 Thai controls. A case-control study showed that the G allele of c.425C>G was significantly associated with ASD (Fisher's exact test, P=0.0359). In addition, the new variant was predicted to be possibly damaging to the EN2 protein by the PolyPhen-2 and FATHMM bioinformatic programs. Our findings suggest that the arginine variant of the EN2 protein may play an important role in the pathology of ASD. Therefore, EN2 protein functional studies should be carried out to determine whether the novel variant has an effect on protein expression.

Objectives: Centrifugation is a time-consuming step which increases the turnaround time (TAT) in laboratories. A few studies have addressed the effect of altering centrifugation settings on analytical quality for clinical chemistry analytes, and most of these studies have used collection tubes with gel separators. However, gel separator tubes may be unsuitable for some laboratories because they are slightly more expensive than tubes without gel separators and are not appropriate for some special tests. The aim of this study was to investigate the effect of centrifugation conditions on clinical chemistry analytes. Methods: We compared centrifugation times of 7 min at 2,200× g and 5 min at 2,750× g with the manufacturer’s protocol of 10 min at 1,300× g as the reference condition. Twenty general chemistry analytes were studied in lithium heparin plasma tubes without gel separators. Results: For all analytes except carbon dioxide (CO ₂ ), no significant differences in analyte results were observed when the centrifugation time was reduced. Deming regression and Bland–Altman plots demonstrated an acceptable clinical concordance within the limits of total allowable error for all analytes between the two rapid centrifugation conditions with the reference centrifugation condition. Conclusions: Our results confirmed that alternate centrifugation conditions for either 7 min at 2,200× g or 5 min at 2,750× g of samples collected in lithium heparin tubes without gel are acceptable for clinical chemistry analytes. Our data support using centrifugation at higher speeds for shorter times to improve TAT without altering the quality of the analytical results.

Fragile X syndrome (FXS) is mainly caused by FMR1 CGG repeat expansions. Other types of mutations, particularly deletions, are also responsible for FXS phenotypes, however these mutations are often missed by routine clinical testing. Molecular diagnosis in cases of suspected FXS was a combination of PCR and Southern blot. Measurement of the FMRP protein level was useful for detecting potentially deleterious impact. PCR analysis and Southern blot revealed a case with premutation and suspected deletion alleles. Sanger sequencing showed that the deletion involved 313 bp upstream of repeats and some parts of CGG repeat tract, leaving transcription start site. FMRP was detected in 5.5 % of blood lymphocytes. According to review of previous cases with FMR1 microdeletions, most patients carrying concurrent deletion and full mutation had typical clinical features of FXS. To our knowledge, our case is the first to describe mosaicism of a premutation and microdeletion in the FMR1 gene. The patient was probably protected from the effects of the deletion by mosaicism with premutation allele, leading to milder phenotype. It is thus important to consider appropriate techniques for detecting FMR1 variants other than repeat expansions which cannot be detected by routine FXS diagnosis.