ABSTRACT
OBJECTIVE@#To analyze gene variants in a Chinese pedigree with oculocutaneous albinism (OCA).@*METHODS@#Gene sequencing of the proband and his parents was performed using chip capture high-throughput sequencing and Sanger sequencing techniques, and PolyPhen-2, SIFT, MutationTaster, and FATHMM software were used to predict the function of new variants. At the same time,the pedigree and variant genes of 4 albinism patients from this pedigree were analyzed.@*RESULTS@#Sequencing results showed that the proband's TYR gene (NM_000372) has c.230G>A (p.Arg77Gln) and c.120_121insG (p.Asp42GlyfsTer35) compound heterozygous variants. The proband's father carries c.230G>A heterozygous variant, and the mother carries c.120_121insG heterozygous variant, indicating that the proband's two variants are from his father and mother. The former is a known missense variant, which can cause abnormal or loss of the original function of the protein polypeptide chain. The latter c.120_121insG(p.Asp42GlyfsTer35) is an unreported frameshift variant of the TYR gene subregion (EX1; CDS1). PolyPhen-2, SIFT, MutationTaster and FATHMM predictions are all prompted as "harmful variants". This variant caused the amino acid encoded protein to terminate prematurely, producing a truncated protein, which eventually formed a 76-amino acid short-type TYR protein instead of the 529-amino acid wild-type TYR protein. Through the pedigree analysis, the four patients in the pedigree are all of the same type of compound heterozygous variants, and the disease-causing genes are all from the patient's parents. They belong to a special form of consanguineous marriage within 5 generations.@*CONCLUSION@#The compound heterozygous variants of c.230G>A (p.Arg77Gln) and c.120_121insG (p.Asp42GlyfsTer35) of the TYR gene may underlie the disease in this pedigree. The gene sequencing results enrich the variant spectrum of the TYR gene, and has facilitated molecular diagnosis for the patient.
Subject(s)
Humans , Albinism, Oculocutaneous/genetics , Consanguinity , Heterozygote , Mutation , PedigreeABSTRACT
Objective To confirm the interference effect of heparin lithium on turbidimetry method for detecting total protein , and to conduct performance evaluation of it .Methods Eight different concentrations of total protein urine specimen with and with ‐out heparin lithium were detected respectively ,to confirm the interference of heparin lithium on the experiment .Adding different concentration of heparin lithium solution to the prepared total protein samples with concentration of 0 .172 ,0 .427 g/L(a series of interference experiment samples ,with heparin lithium concentrations of 0 .000 ,0 .025 ,0 .030 ,0 .035 ,0 .040 ,0 .045 ,0 .050 mg/L ) , repeat four times detection ,and then conduct the dose‐effect experiment of interference .Results The biggest bias of the testing re‐sults in the two paired samples was 159 .37% ,the mini‐bias was 11 .37% ,the difference of the results in two groups was statistical significant(t= 17 .24 ,P< 0 .05) .95% confidence interval of the interference effect in the total protein samples with concentration of 0 .172 g/L was 0 .034 ~ 0 .062 ,while in the total protein samples with concentration of 0 .427 g/L was 0 .043 - 0 .053 .When the heparin lithium were 0 .025 mg/mL and 0 .050 mg/mL respectively ,the results of bias of the total protein samples with concentra‐tion of 0 .172 g/L were 27 .33% ,58 .14% respectively .But in the 0 .427 g/L total protein samples ,the results of bias were 11 .24% 、18 .74% respectively .Dose effect of interference both showed a linear relationship in the series concentration of the hepa ‐rin lithium and interference of sample of 0 .172 g/L and 0 .427 g/L ,and the linear equations were Y = 1 .974X - 0 .001 03 ,Y = 1 .599 X - 0 .000 5 respectively .Conclusion Heparin lithium is the exogenous interfering substance to the detection of total protein in tur ‐bidimetry method ,it is the positive interference to the determination results of total proteins ,especially for the low level of total pro‐tein detection ,and as heparin lithium concentration increases ,the stronger the interference effect .
ABSTRACT
Objective To compare the results difference between the fluorescence staining and the acid fast staining (Ziehi‐Neelsen staining) methods in the detection of Mycobacterium tuberculosis ,and to compare the effects of the methylene blue solu‐tion ,Haris hematoxylin solution and potassium permanganate liquid as the redyeing reagents of the fluorescence staining method . Methods 198 sputum specimens collected from the patients with suspected tuberculosis symptoms and were performed the Ziehi‐Neelsen staining and the fluorescence staining respectively For comparing the difference in the detecting rate of Mycobacterium tu‐berculosis between the two kinds of method .The fluorescence staining adopted 0 .3% methylene blue solution ,0 .5% Haris hema‐toxylin solution and 0 .5% potassium permanganate solution as the redyeing reagents for comparing the effects of the fluorescence microscopic examination among different redying reagents .Results The detection rate of Mycobacterium tuberculosis was 66 .67%(132/198) for the Ziehi‐Neelsen staining ,94 .9% (188/198) for the fluorescence stainings and 94 .95% (188/198) for the methyl‐ene blue staining ,in which the detection rate of methylene blue redyeing was 94 .95% ,which of hematoxylin redyeing was 94 .44%(187/198) and which of potassium permanganate redyeing was 94 .44 (187/198) ,the differences among them were statistically sig‐nificant(P< 0 .05) .Conclusion The fluorescent staining method has the higher positive detection rate of Mycobacterium tubercu‐losis than the Ziehl‐Neelsen staining method ,in which 0 .3% methylene blue solution is a good background quenching redyeing solu‐tion .