CAJ | 학술논문

目的比较串联质谱两种前标本处理方法(衍生化法和非衍生化法)检测新生儿滤纸干血片上丙氨酸(Ala),精氨酸(Arg),瓜氨酸(Cit),甘氨酸(Gly),亮氨酸(Leu),甲硫氨酸(Met),鸟氨酸(Orn),苯丙氨酸(Phe),酪氨酸(Tyr),缬氨酸(Val) 10种氨基酸水平分布和差异,为实验室开展新生儿氨基酸代谢障碍疾病筛查提供方法学依据.方法方法比较研究.选择2012年1至6月在上海市儿童医院新生儿筛查中心进行遗传代谢性疾病筛查滤纸干血片标本,共计4135份.采用衍生化法和非衍生化法及相应试剂盒:(1)同时检测100份新生儿滤纸干血片标本,以配对方式比较10种氨基酸水平之间差异；(2)分别各自检测2000份正常新生儿滤纸干血片标本,了解正常新生儿10种氨基酸水平分布；(3)同时检测经临床确诊的7种氨基酸代谢异常标本35份,验证两种方法临床判断的一致性.采用配对t检验.结果正常新生儿的10种氨基酸水平经K-S单标本正态分布检验,Z值在1.997 ～6.229之间,呈偏态分布(P＜0.01)；除Leu和Tyr外,其余8种氨基酸非衍法比衍生法检测浓度低,两种方法检测均值差异,除Met高达47.8％外,其他9种氨基酸水平差异均＜10％,平均为7.8％；除Met、Phe和Tyr,两种方法测得的7种氨基酸水平差异无统计学意义(P＞0.05)；以0.5～99.5百分位数建立的10种氨基酸正常参考值范围,衍生法均比非衍法高,平均高25.3％,正常参考值范围有重叠；经临床验证,对于确诊患儿异常指标的检测,两种方法间无统计学意义(P＞0.05),两种方法均能检测出疾病患儿,检出率均为100％.结论衍生法和非衍法检测多种氨基酸水平有差异,但对氨基酸代谢障碍疾病的异常指标的检测结果无差异,对疾病的临床判断无影响；两种方法均可用于新生儿氨基酸代谢障碍疾病的筛查.
목적 비교천련질보량충전표본처리방법(연생화법화비연생화법)검측신생인려지간혈편상병안산(Ala),정안산(Arg),과안산(Cit),감안산(Gly),량안산(Leu),갑류안산(Met),조안산(Orn),분병안산(Phe),락안산(Tyr),힐안산(Val) 10충안기산수평분포화차이,위실험실개전신생인안기산대사장애질병사사제공방법학의거.방법 방법비교연구.선택2012년1지6월재상해시인동의원신생인사사중심진행유전대사성질병사사려지간혈편표본,공계4135빈.채용연생화법화비연생화법급상응시제합:(1)동시검측100빈신생인려지간혈편표본,이배대방식비교10충안기산수평지간차이；(2)분별각자검측2000빈정상신생인려지간혈편표본,료해정상신생인10충안기산수평분포；(3)동시검측경림상학진적7충안기산대사이상표본35빈,험증량충방법림상판단적일치성.채용배대t검험.결과 정상신생인적10충안기산수평경K-S단표본정태분포검험,Z치재1.997 ～6.229지간,정편태분포(P＜0.01)；제Leu화Tyr외,기여8충안기산비연법비연생법검측농도저,량충방법검측균치차이,제Met고체47.8％외,기타9충안기산수평차이균＜10％,평균위7.8％；제Met、Phe화Tyr,량충방법측득적7충안기산수평차이무통계학의의(P＞0.05)；이0.5～99.5백분위수건립적10충안기산정상삼고치범위,연생법균비비연법고,평균고25.3％,정상삼고치범위유중첩；경림상험증,대우학진환인이상지표적검측,량충방법간무통계학의의(P＞0.05),량충방법균능검측출질병환인,검출솔균위100％.결론 연생법화비연법검측다충안기산수평유차이,단대안기산대사장애질병적이상지표적검측결과무차이,대질병적림상판단무영향；량충방법균가용우신생인안기산대사장애질병적사사.
Objective To provide methodological references for laboratories to carry out newborn screening for disorders of amino acid metabolism,we compared the difference and distribution of ten amino acids including alanine (Ala),arginine (Arg),citrulline (Cit),glycine (Gly),leucine (Leu),methionine (Met),ornithine (Orn),phenylalanine (Phe),tyrosine (Tyr),and valine (Val) from newborn dried blood spots specimen using derivatization or non-derivatization as sample preparation methods.Method It is a comparative research study.A total of 4135 newborn screening dried blood spot samples for inborn errors of metabolism were collected from January to June,2012.All specimens came from neonatal screening center of shanghai children's hospital.Samples were prepared by two different techniques,the corresponding kits and the procedures were used as follows:(1) Simultaneous detection of 100 dried blood spot specimens using two methods respectively to compare the paired difference of each amino acid.(2) 2000 cases of normal newborn specimens were detected respectively to obtain the normal distribution of ten neonatal amino acids.(3) 35 specimens from patients previously diagnosed positively as inborn errors of metabolism were simultaneously detected with 7 amino acids to verify the consistency of two techniques in clinical judgment.Results The amino acid levels of normal newborns analyzed by one-sample.kolmogorovSmirnov test (Z value ranged from 1.997 to 6.229) showed a skewed distribution (P ＜ 0.01).Except for Leu and Tyr,non-derivatization techniqueshowed a lower concentration than derivatization technique,and the CVs of nine amino acids were ＜ 10％ except for Met (the CV of Met was 47.8％),and the average CV is 7.8％.Except for Met,Phe and Tyr,the levels of other 7 amino acids measured by two techniques showed no significant difference (P ＞ 0.05).According to 0.5th to 99.5th percentiles,the normal reference range for derivatization method were greater than on-derivatization method,and the average value was 25.3％.After clinic judgment,the results of the abnormal indicators of children with true metabolism disorders showed no statistically significant between two methods (P ＞ 0.05),the detection rate was 100％.Conclusions There was a slight difference between derivatization and the non-derivatization techniques in detecting multiple amino acids.The results of the abnormal indicators of amino acid metabolism disordersshow no statically significant difference between the two methods,and no difference in clinical judgment.Both methods can be used in detecting amino acid metabolism disorders in newborn screening.