SMN1 gene deletion analysis using mid-trimester amniotic fluid cells by real-time PCR / 中华检验医学杂志

Objectives To investigate the prenatal diagnosis method of spinal muscular atrophy with amniotic fluid sample.Methods Totally 1 064 amniotic fluid samples from mid-trimester pregnant women were enrolled during January 2015 and January 2016 in 4 hospitals.Genetic analysis was performed for detecting potential contamination of maternal tissue by a genetic technique based on short tandem repeat ( STR) markers.Deletion of SMN1 gene was detected in 1 062 uncontaminated amniotic fluid samples by real-time PCR and multiplex ligation-dependent probe amplification ( MLPA) respectively.Results Two contaminated amniotic fluid samples were detected within 1 064 mid-trimester pregnant women by STR genotyping.The other 1 062 uncontaminated amniotic fluid samples were tested by real-time PCR.There were 37 samples with heterozygous deletion of Exon 7 of SMN1 gene ( 3.67%) , 34 samples with heterozygous deletion of Exon 8 of SMN1 gene (3.2%) and two samples with homozygous deletion of Exon 7 and Exon8 of SMN1 gene ( 0.19%) respectively , while other samples observed with no deletion of Exon 7 and Exon8 in SMN1 gene.Totally 41 samples with heterozygous or homozygous deletion of SMN 1 gene and 55 samples with undetected deletion of SMN 1 gene were confirmed by MLPA and the results showed 100%consistence with that of real-time PCR.Conclusions Both real-time PCR and MLPA are suitable for detecting the deletion of SMN 1 gene with amniotic fluid sample . Real-time PCR exhibits less sample requirement and time compared with MLPA .

Clinical significance of HPV subtype infection and atypical glandular cells on LBP / 国际检验医学杂志

Objective To research the clinical outcome of atypical glandular cell (AGC)according to various subtypes of HPV infection and histological pathology results.Methods The data of the liquid-bases cytology (LBP),HPV infection and histology in 102 cases of AGC at the gynecology outpatient department of our hospital from January 1 ,2009 to February 28,2014 were collected and performed the analysis on their clinical outcomes.Results Among 67 218 cases of LBP detection,102 cases were AGC with the total incidence rate of 0.15%.In the cases of AGC-NOS,67 cases were normal or benign lesions,11 cases were precancerous lesion and malignant lesions;in the cases of AGC treading to tumor,the benign,precancerous and malignant lesions were in 7,14 and 3 ca-ses respectively.At the same time in the cases of AGC-NOS,HPV infection was in 64 cases,in which 57 cases were high risk infec-tion(type 16,52,45)and 7 cases were low risk infection(type 6,11 ).The single infection,double infection and multiple infection were in 54,6 cases respectively;in the cases of AGC trends to neoplasm,HPV infection was in 19 cases,in which 18 cases were high risk infection(type 52,16,18)and 1 case was low risk infection(type 6),single infection and double infection were in 15 cases and 4 cases respectively.Conclusion AGC may play an important role for the forecast of cervical malignant lesions.The results of differ-ent HPV subtypes infection in AGC related tumors also play a certain role in the prediction of cervical neoplasia.Their combined a-nalysis is the important signal for judging the occurrence of gynecological cervical precancerous lesion and malignant tumor,i.e., AGC combining with the corresponding HPV subtype infection not only can make a judgement for the cervical lesions,but also pro-vides the basis for predicting the high risk existence of gynecological malignant tumor and provides constructive suggestions for Chi-na regional cervical carcinoma vaccine manufacturing and promotion.

The application of alpha-fetoprotein isoform L2 test in screening Down syndrome / 国际检验医学杂志

Objective To explore significance of alpha‐fetoprotein isoform L2(AFP‐L2) in the screening of Down syndrome in pregnant women ,so as to provide references for clinical application .Methods A total of 250 healthy pregnant women and 22 preg‐nant women with Down syndrome were enrolled in this study .Serum specimens were collected and AFP‐12 was separated and cap‐tured by using the magnetic bal ,time‐resolved fluorescence immunoassay was used to detect levels of AFP and AFP‐L2 ,and the percentage of AFP‐L2 (AFP‐L2% ) was calculated .Results The serum level of AFP of pregnant women with Down syndrome [(20.2±4.2)ng/mL]was lower than that of healthy pregnant women[(46.7±19.9)ng/mL],and had statistically significant difference(P<0 .05) .Serum AFP‐L2% of pregnant women with Down syndrome was higher than that of healthy pregnant women , and had statistically significant difference(P<0 .05) .Conclusion Detection of AFP level and AFP‐L2% could be an indicator for Dow n syndrome screening .

CK assay with gene testing for screening of Duchenne muscular dystrophy in the newborn infants / 中华检验医学杂志

Objective To establish a system using CK activity assessing with follow-up Duchenne muscular dystrophy (DMD) gene testing to newborn screening for DMD.This study provided a pathway to improve the health outcome for individuals with DMD.Methods Tests for CK were performed with Beckman original reagent on a Beckman Coulter AU 5800.Preliminary studies established a population-based range of CK in newborns using 5 892 deidentified anonymous blood samples,which were collected from Shanghai Changning Maternity and Infant Health Hospital between November 2013 and July 2014.Mutation analysis used multiplex PCR-denature high-performance liquid chromatography (PCR-DHPLC) method for screening large duplications and deletions and Sanger DNA sequencing for screening point DMD gene mutation.Results DMD gene mutations (point mutation,exon60,c.9072G ＞ A) were found in 1 of 5 892 newborn subjects,which had CK level ＞ 2 000 U/L large duplications and deletions in DMD gene were not found.Conclusions A system of analysis for newborn screening for DMD has been established.This path for newborn screening fits our health care system and minimizes the false-positive results for predicting DMD gene mutations by use of CK levels in blood