早孕期血清PAPP-A檢測聯(lián)合中孕期血清AFP、free β-HCG、uE3檢測對DS、ES的篩查效果

楊嵐,石錦平,吳曉,施楠,郭彩琴,趙麗

(南京醫(yī)科大學(xué)附屬無錫婦幼保健院, 江蘇無錫214002)

早孕期血清PAPP-A檢測聯(lián)合中孕期血清AFP、free β-HCG、uE3檢測對DS、ES的篩查效果

楊嵐,石錦平,吳曉,施楠,郭彩琴,趙麗

(南京醫(yī)科大學(xué)附屬無錫婦幼保健院,江蘇無錫214002)

目的觀察早孕期檢測血清PAPP-A聯(lián)合中孕期檢測血清AFP、free β-HCG、uE3對胎兒唐氏綜合征(DS)和愛德華氏綜合征(ES)的篩查效果。方法回顧性分析在我院行產(chǎn)前篩查且有隨訪結(jié)局的7 055例孕婦的臨床資料。所有孕婦早孕期檢測血清妊娠相關(guān)蛋白A(PAPP-A)和游離人絨毛膜促性腺激素β(Free β-hCG)，中孕期檢測血清甲胎蛋白(AFP)、free β-HCG、游離雌三醇(μE3)。設(shè)計(jì)3個(gè)DS、ES篩查方案。方案A納入指標(biāo)為早孕期PAPP-A、Free β-hCG，方案B案納入指標(biāo)為中孕期AFP、free β-HCG、μE3，方案C納入指標(biāo)為早孕期PAPP-A和中孕期AFP、free β-HCG、μE3。用LifeCycle 4.0軟件綜合患者年齡、體質(zhì)量、孕周等數(shù)據(jù)計(jì)算用三種方案篩查得出的患者DS、ES風(fēng)險(xiǎn)值。DS風(fēng)險(xiǎn)值>1/270為高風(fēng)險(xiǎn)，1/1 000～1/270為臨界風(fēng)險(xiǎn)；ES風(fēng)險(xiǎn)值>1/350為高風(fēng)險(xiǎn)，1/1 000～1/350為臨界風(fēng)險(xiǎn)。比較三種方案的DS高風(fēng)險(xiǎn)、DS臨界風(fēng)險(xiǎn)、ES高風(fēng)險(xiǎn)、ES臨界風(fēng)險(xiǎn)及陽性篩查結(jié)果檢出情況。胎兒出生者以胎兒是否確診DS、ES為金標(biāo)準(zhǔn)，終止妊娠者以羊水核型分析結(jié)果為金標(biāo)準(zhǔn)，計(jì)算三種方案篩查DS、ES的檢出率、假陽性率。結(jié)果方案C的DS高風(fēng)險(xiǎn)、DS臨界風(fēng)險(xiǎn)、篩查結(jié)果總陽性率均低于方案A、B(P均<0.05)。羊水穿刺染色體核型分析和胎兒出生后隨訪共發(fā)現(xiàn)DS患兒9例，其中方案A篩查結(jié)果陽性7例，檢出率77.8%，假陽性率3.95%；方案B篩查結(jié)果陽性6例，檢出率66.7%，DS假陽性率4.24%；方案C篩查結(jié)果陽性8例，檢出率88.9%，假陽性率1.80%。方案C的檢出率、假陽性率均優(yōu)于方案A、B。羊水穿刺染色體核型分析和胎兒出生后隨訪共發(fā)現(xiàn)ES患兒2例，其中1例在3種篩查方案都得到了檢出，另1例ES患者用聯(lián)合方案得出了高風(fēng)險(xiǎn)值，ES檢出率聯(lián)合篩查方案均高于孕早期、孕中期方案。結(jié)論早孕期檢測血清PAPP-A聯(lián)合中孕期檢測血清AFP、free β-HCG、uE3對胎兒DS和ES的篩查效果優(yōu)于早孕期檢測血清PAPP-A、free β-HCG或中孕期檢測血清AFP、free β-HCG、uE3。

唐氏綜合征；愛德華氏綜合征；早孕期；中孕期；妊娠相關(guān)蛋白A；人絨毛膜促性腺激素β；甲胎蛋白；雌三醇

Abstract:ObjectiveTo explore the screening efficiency about the first trimester screening of pregnancy-associated plasma protein A (PAPP-A) combined with the second trimester screening of alpha fetoprotein (AFP), free β-human chorionic gonadotropin (free β-HCG), uncojugated estriol (uE3) strategies for Down's Syndrome (DS) and Edwards' syndrome (ES).MethodsThe clinical data from 7 055 cases of pregnant women with pregnancy outcome, who underwent prenatal screening test, were retrospectively analyzed. The serum PAPP-A, free β-hCG in the first trimester, and AFP, free β-HCG, and uE3 in the second trimester were detected in all pregnant women. We designed three screening tests as three groups: projects A, B, and C. The risk evaluation of project A was serum free β-hCG and PAPP-A levels in the first trimester screening. Project B included free β-hCG, AFP, and uE3 levels in the second trimester. As for project C-so called the integrated serum test, included the serum PAPP-A levels in the first trimester, and AFP, β-hCG, and uE3 in the second trimester. The risks of DS and ES were evaluated by LifeCycle 4.0 software based on the age, weight and gestational week of gravidas. The cut-off value (high risk) and the intermediate risk value were >1/270 and 1/1 000-1/270 in DS, >1/350 and 1/1000-1/350 in ES, respectively. We compared the high risk rate, intermediate risk rate in both DS and ES as well as the total positive rates among the three screening tests mentioned above. The women who terminated pregnancy were identified by prenatal diagnosis via amniocentesis. Furthermore, those who did not terminate pregnancy were confirmed by the pregnancy outcomes, and were followed up based on karyotype of newborns. We evaluated screening efficiency of DS and ES such as the detection rate (DR) and false positive rate (FPR) among the three screening strategies.ResultsThe high risk and the intermediate risk in DS as well as the screening positive rate of the project C were significant lower than those of projects A and B, respectively (P<0.05) (neural tube defect were excluded). A total of 9 fetus with true DS were diagnosed via amniocentesis and follow-up pregnancy outcomes. In the project A, the number of high risk was 7, DR and FPR for trisomy 21 was 77.8% (7/9) and 3.95% (279/7 055), respectively. In the project B, the number of high risk was 6, DR and FPR for trisomy 21 was 66.7% (6/9) and 4. 24% (299/7 055), respectively. In the project C, DR and FPR for trisomy 21 was 88.9% (8/9) and 1.80% (127/7 055) , respectively. The DR and FPR for DS in the project C were both superior to those in the projects A and B (DR:P>0.05, FPR:P<0.05). We diagnosed two fetus with true ES through amniocentesis and follow-up pregnancy outcomes. Only one case was successfully detected by all three screening projects. The other case was detected as high risk in the combined screening test. The detection rate for ES in the project C was higher than those in both projects A and B.ConclusionThe screening efficiency about the first trimester screening of PAPP-A combined with the second trimester screening of AFP, free β-HCG, and uE3 for DS and ES is superior to both the first trimester double test (PAPP-A and free β-HCG) and the second trimester triple test including AFP, free β-HCG, and uE3.

Keywords: Down Syndrome; Edwards syndrome; the first trimester; the second trimester; pregnancy-associated plasma protein A; human chorionic gonadotropin-β; alpha fetoprotein; estriol

唐氏綜合征(DS，又名21三體綜合征)和愛德華氏綜合征(ES，又名18三體綜合征)是臨床常見的先天出生缺陷病。針對這類胎兒染色體疾病的產(chǎn)前篩查方案，現(xiàn)今國內(nèi)應(yīng)用最普遍的是中孕期血清學(xué)指標(biāo)篩查方案[1～2]，該方法經(jīng)濟(jì)簡便，對胎兒無創(chuàng)，可在一定程度上減少這類缺陷兒出生[3]，但其檢出率較低[4～5]。國外有研究顯示將早、中孕期血清學(xué)指標(biāo)整合為一個(gè)方案，可將DS、ES檢出率提高到88%～95%[6，7]。但國內(nèi)尚無同類文獻(xiàn)報(bào)道。本研究對2015年5月～2016年5月在我院規(guī)律產(chǎn)檢的7 055例單胎孕婦的臨床資料進(jìn)行了回顧行分析，觀察早孕期檢測血清PAPP-A聯(lián)合中孕期檢測血清AFP、free β-HCG、uE3(聯(lián)合方案)對胎兒DS和ES的篩查效果。現(xiàn)報(bào)告如下。

1 資料與方法

1.1 臨床資料 選擇同期于我院產(chǎn)前診斷中心規(guī)律產(chǎn)檢且有完整妊娠結(jié)局隨訪結(jié)果的單胎妊娠孕婦共7 055例，預(yù)產(chǎn)年齡8～9歲(28.38±2.95歲)。均于早孕期(孕10～13+6周)和中孕期(15～20+6周)進(jìn)行了產(chǎn)前血清學(xué)指標(biāo)篩查。均無吸煙史，無胰島素依賴性糖尿病，無DS、ES兒分娩史。本研究已經(jīng)我院醫(yī)學(xué)倫理委員會批準(zhǔn)。孕婦或其家屬均簽署知情同意書，并填寫早、中孕期產(chǎn)前篩查申請單。

1.2 DS、ES篩查方法 ①方案A：孕10～13+6周抽取孕婦肘靜脈血3 mL，不抗凝，室溫靜置2 h后于3 500 r/min離心6 min后分離血清，-20 ℃儲存?zhèn)錂z。采用 芬蘭PerKin Elmer公司生產(chǎn)的1235全自動多標(biāo)記儀、DELFIA時(shí)間分辨熒光法及配套試劑檢測血清妊娠相關(guān)蛋白A(PAPP-A)和游離人絨毛膜促性腺激素β(free β-hCG)水平。操作按試劑盒說明書進(jìn)行。用Lifecycler 4.0軟件綜合患者年齡、體質(zhì)量、孕周等數(shù)據(jù)計(jì)算患者用早孕期方案篩查DS、ES風(fēng)險(xiǎn)值。DS風(fēng)險(xiǎn)值>1/270為高風(fēng)險(xiǎn)，風(fēng)險(xiǎn)值1/1 000～1/270為臨界風(fēng)險(xiǎn)；ES風(fēng)險(xiǎn)值>1/350為高風(fēng)險(xiǎn)，1/1 000～1/350為臨界風(fēng)險(xiǎn)。②方案B：納入的血清學(xué)指標(biāo)為孕中期(孕15～20+6周)血清甲胎蛋白(AFP)、free β-hCG和游離雌三醇(uE3)。其余同孕早期方案。③方案C：納入的血清學(xué)指標(biāo)為孕中期血清AFP、free β-hCG、uE3和孕早期PAPP-A。其余同孕早期方案。

1.3 DS、ES篩查的準(zhǔn)確性評價(jià)方法 對篩查出的高風(fēng)險(xiǎn)孕婦進(jìn)行遺傳咨詢，建議其接受羊水穿刺進(jìn)行胎兒染色體核型分析。孕婦胎兒出生者均進(jìn)行隨訪確定胎兒是否患有DS或ES。胎兒出生者以胎兒是否確診DS、ES為金標(biāo)準(zhǔn)，終止妊娠者以羊水核型分析結(jié)果為金標(biāo)準(zhǔn)，計(jì)算三種方案篩查DS、ES的檢出率、假陽性率。

1.4 統(tǒng)計(jì)學(xué)方法 采用SPSS17.0統(tǒng)計(jì)軟件。率的比較采用χ2檢驗(yàn)和Fisher確切概率法。P<0.05 為差異有統(tǒng)計(jì)學(xué)意義。

2 結(jié)果

2.1 三種方案篩查結(jié)果比較 三種方案篩查結(jié)果見表1。方案C的DS高風(fēng)險(xiǎn)、DS臨界風(fēng)險(xiǎn)率均低于方案A和B(P均<0.05)，而其ES高風(fēng)險(xiǎn)、ES臨界風(fēng)險(xiǎn)率無統(tǒng)計(jì)學(xué)差異(P均>0.05)；方案A和B的DS高風(fēng)險(xiǎn)、ES高風(fēng)險(xiǎn)、ES臨界風(fēng)險(xiǎn)率相比，P均>0.05。

表1 三種方案篩查結(jié)果[例(%)]

2.2 三種方案篩查結(jié)果準(zhǔn)確性比較 羊水穿刺染色體核型分析和胎兒出生后隨訪共發(fā)現(xiàn)DS患兒9例(其中1例為易位型)，其中方案A篩查結(jié)果陽性7例，靈敏度77.8%，假陽性率3.95%；方案B篩查結(jié)果陽性6例，靈敏度66.7%，假陽性率4.24%；方案C篩查結(jié)果陽性8例，假陽性率1.80%。方案C的檢出率均高于方案A和B(P均<0.05)，假陽性率均低于方案A和B(P均<0.05)。見表2。

表2 三種方案篩查DS效果比較

2.3 不同篩查方案對11例胎兒非整倍體疾病的篩查風(fēng)險(xiǎn)值比較 詳見表1方案C篩查得出的DS、ES兒風(fēng)險(xiǎn)值均比方案A和B高。

3 討論

前已述及，現(xiàn)今產(chǎn)前篩查的各種篩查方案都有不同的檢出率和假陽性率，檢出率>80%且假陽性率<5%是比較理想的結(jié)果[8]。本研究結(jié)果顯示三種篩查方案的特異度都>95%，方案C的特異度最高；方案A、B、C篩查DS檢出率分別為77.8%、66.7%、88.9%，方案C的檢出率高于方案A、B。另外本研究方案B的DS檢出率與國內(nèi)外報(bào)道相符[1，2]，方案A的檢出率略低于文獻(xiàn)報(bào)道的85%[4，5]，分析可能與未納入超聲指標(biāo)頸項(xiàng)透明層厚度(NT)有關(guān)，研究顯示，如納入NT指標(biāo)則有望將DS的檢出率提高到89%以上[9]。早孕篩查最大的優(yōu)勢在于可使高風(fēng)險(xiǎn)孕婦在孕早期即通過絨毛染色體分析得到確診，以使真陽性孕婦及時(shí)終止妊娠，是目前發(fā)達(dá)國家較常用的篩查方案[10]。本研究方案A、B的DS高風(fēng)險(xiǎn)、DS臨界風(fēng)險(xiǎn)及總陽性結(jié)果檢出雖然比方案C多，但是檢出率低于方案C，假陽性率則高于方案C，與相關(guān)報(bào)道[11，12]相仿。

有研究顯示酌情篩查和早中孕全面整合篩查的假陽性率僅2%～3%，DS檢出率達(dá)到90%以上[7, 13]。但全面整合篩查需符合NT檢查要求，酌情篩查需將孕婦合理區(qū)分為高風(fēng)險(xiǎn)、臨界風(fēng)險(xiǎn)、低風(fēng)險(xiǎn)三類，雖然在篩查成本上較整合篩查更優(yōu)[14]，但需確定該方案的各類風(fēng)險(xiǎn)范圍，不同實(shí)驗(yàn)室不同地區(qū)的標(biāo)準(zhǔn)并不相同。此外，該方案和序貫篩查一樣，在早孕期的高風(fēng)險(xiǎn)孕婦需行絨毛穿刺加以確診，故限制了這兩類方案的廣泛應(yīng)用，而整合篩查方案既可獲得較高的檢出率，又減少了經(jīng)濟(jì)壓力和有創(chuàng)診斷的社會支出，在早孕期還緩解了孕婦的焦慮情緒，具備較好的成本效益值，在NT檢查和絨毛穿刺技術(shù)受局限地區(qū)，適于臨床推廣。本研究中確診的2例ES患兒，用方案A、B篩查僅有1例得到了高風(fēng)險(xiǎn)值，另1例經(jīng)方案C篩查判定為高風(fēng)險(xiǎn)，可見方案C篩查較方案A、B增加了1例ES的檢出。并且方案A、B篩查出的1例ES患兒用方案C測算出的ES風(fēng)險(xiǎn)值高于方案A和B，體現(xiàn)出了方案C對ES良好的篩查價(jià)值，該優(yōu)勢在國外學(xué)者的研究中也得以證實(shí)[15]。

本研究結(jié)果顯示，方案C可以減少陽性篩查結(jié)果數(shù)量(假陽性率)，從而減少羊水穿刺這種有創(chuàng)性檢查的數(shù)量。雖然臨床上還有一種敏感且無創(chuàng)的高通量測序技術(shù)通過對母血中的胎兒游離DNA進(jìn)行測序?qū)S、ES進(jìn)行篩查[16]，但目前其高昂的檢測成本限制了其普遍開展。本研究結(jié)果顯示 早孕期檢測血清PAPP-A聯(lián)合中孕期檢測血清AFP、free β-HCG、uE3對DS和ES體現(xiàn)出了良好的篩查效率，有助于減低患者經(jīng)濟(jì)壓力和社會支出，性價(jià)比較高，適于臨床推廣。

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Screening efficiency about the first trimester screening of PAPP-A combined with the second trimester screening of AFP, free β-HCG, and uE3 strategies for Down's Syndrome and Edwards' syndrome

YANGLan,SHIJingping,WUXiao,SHINan,GUOCaiqin,ZHAOLi

(WuxiMaternalandChildHealthHospitalAffiliatedtoNanjingMedicalUniversity,Wuxi214002,China)

10.3969/j.issn.1002-266X.2017.35.006

R714.7

A

1002-266X(2017)35-0018-04

2017-03-14)

無錫市醫(yī)管中心面上項(xiàng)目(YGZXM1510)；無錫市科技發(fā)展基金項(xiàng)目(CSE31N1511)；江蘇省婦幼保健重點(diǎn)資助項(xiàng)目(201315)。

楊嵐(1979-),女，醫(yī)學(xué)碩士，副主任醫(yī)師, 主要研究方向?yàn)楫a(chǎn)前篩查、產(chǎn)前診斷。E-mail：lilylan5930@sina.com

趙麗(1978-)，女，醫(yī)學(xué)碩士，副主任醫(yī)師，主要研究方向?yàn)楫a(chǎn)前診斷、遺傳咨詢。E-mail：our163@163.com