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滤纸采集外周血对新生儿进行特异性IgA和IgM抗体的血清学筛查。

Serological Screening of Newborns for -Specific IgA and IgM Antibodies in Peripheral Blood Collected on Filter-Papers.

作者信息

Paul Malgorzata

机构信息

Department and Clinic of Tropical and Parasitic Diseases University of Medical Sciences, Poznan, Poland.

出版信息

EJIFCC. 2007 Oct 4;18(3):91-114. eCollection 2007 Oct.

Abstract

UNLABELLED

The strategic approach for preventing congenital toxoplasmosis is strictly related to the incidence of primary infection during pregnancy in a studied population. Early postnatal diagnosis by mass testing of newborns is an option in areas where obligatory serological screening in pregnant women has not been implemented but it requires sensitive immunodiagnostic methods followed by a good confirmatory analysis. The aims of the regional neonatal screening programme were (i) analysis of the prevalence of congenital infection at birth in the West Poland Province, (ii) determination of the value of the serological examination of filter-paper blood specimens collected at birth for the diagnosis of congenital toxoplasmosis, and (iii) evaluation of the duration of -specific immunoglobulin A and immunoglobulin M antibodies in infants' sera. The neonates born in the obstetric clinics of the University Gynaecology-Obstetrics Hospital in Poznan (Poland) and in the maternity wards of the 10 main district hospitals from the West Poland region were systematically screened for congenital infection. Peripheral blood from newborns was collected by a non-invasive heel-stick puncture during the first 3 days of life, absorbed onto Guthrie cards and analysed for anti- specific IgM (1996-1998) or both IgA and IgM antibodies (1998-2000) by non-commercial immunocapture ELISAs. When the screening result was positive, the diagnosis of congenital infection was confirmed by testing serum samples from the suspected neonate and the mother using a Western blot IgM-IgG comparative immunological profile analysis and traditional serological techniques (ELISA, ISAGA) for anti- IgA, IgM and IgG specific antibodies. From June 1996 to April 2000, 45,169 filter-paper specimens from liveborn neonates were screened: 27,516 samples were tested for specific IgM and the next 17,653 Guthrie cards were analysed by the combined IgA/IgM assay. The prevalence of anti- IgM in filter-paper eluates at birth was 1 per 2,117 liveborn neonates (0.47/1000) or 1 per 1,185 infants (0.84/1000) born to seronegative women with a potential risk of primary infection during pregnancy. For the joint detection of IgA and IgM, these values significantly increased to 1 per 929 neonates (1.08/1000) or 1 per 520 pregnancies at risk (1.92/1000) respectively, comparing to the seropositivity rate of 43.7% in a pregnant women population in the studied area. In newborns untreated prenatally, the diagnostic sensitivity of the IgM ELISA using neonatal Guthrie cards was not more than 86.7% and that of the combined IgA/IgM ELISA was 95%; the diagnostic specificity of the both methods was calculated to be 99.9%. Congenital infection was finally diagnosed in 35 neonates, mostly asymptomatic at birth.

CONCLUSIONS

(i) The neonatal screening for anti- IgA and/or IgM antibodies is a good sensitivity method for an early postnatal diagnosis of congenital toxoplasmosis in newborns untreated prenatally. (ii) In the absence of obligatory nation-wide screening during pregnancy followed by an early prenatal treatment, this valuable technique may be considered a preventive option in areas of a high annual number of births associated with a high seroprevalence of infection.

摘要

未标注

预防先天性弓形虫病的策略方法与所研究人群中孕期原发性感染的发生率密切相关。在未实施孕妇强制性血清学筛查的地区,通过对新生儿进行大规模检测进行产后早期诊断是一种选择,但这需要灵敏的免疫诊断方法,随后进行良好的确诊分析。该地区新生儿筛查项目的目的是:(i)分析波兰西部省份出生时先天性感染的患病率;(ii)确定出生时采集的滤纸血标本血清学检查对先天性弓形虫病诊断的价值;(iii)评估婴儿血清中特异性免疫球蛋白A和免疫球蛋白M抗体的持续时间。对在波兹南(波兰)大学妇产科医院产科诊所出生的新生儿以及波兰西部地区10家主要地区医院产科病房出生的新生儿进行先天性感染的系统筛查。在出生后的前3天,通过无创足跟穿刺采集新生儿外周血,吸附在格思里卡片上,并通过非商业免疫捕获酶联免疫吸附测定法分析抗特异性IgM(1996 - 1998年)或IgA和IgM抗体(1998 - 2000年)。当筛查结果为阳性时,通过使用蛋白质印迹IgM - IgG比较免疫图谱分析以及针对抗IgA、IgM和IgG特异性抗体的传统血清学技术(酶联免疫吸附测定法、间接血凝抑制试验)检测疑似新生儿和母亲的血清样本,来确诊先天性感染。从1996年6月到2000年4月,对45169份活产新生儿的滤纸标本进行了筛查:27516份样本检测了特异性IgM,接下来的17653张格思里卡片通过联合IgA/IgM检测进行分析。出生时滤纸洗脱液中抗IgM的患病率为每2117名活产新生儿中有1例(0.47/1000),或每1185名有孕期原发性感染潜在风险的血清阴性妇女所生婴儿中有1例(0.84/1000)。对于IgA和IgM的联合检测,这些值分别显著增加到每929名新生儿中有1例(1.08/1000)或每520例有风险的妊娠中有1例(1.92/1000),而所研究地区孕妇人群的血清阳性率为43.7%。在未进行产前治疗的新生儿中,使用新生儿格思里卡片的IgM酶联免疫吸附测定法的诊断敏感性不超过86.7%,联合IgA/IgM酶联免疫吸附测定法的诊断敏感性为95%;两种方法的诊断特异性经计算均为99.9%。最终确诊35例先天性感染新生儿,大多数出生时无症状。

结论

(i)对新生儿进行抗IgA和/或IgM抗体的筛查是对未进行产前治疗的新生儿先天性弓形虫病进行产后早期诊断的一种敏感性良好的方法。(ii)在没有全国性孕期强制性筛查及早期产前治疗的情况下,对于每年出生人数众多且弓形虫感染血清阳性率高的地区,这项有价值的技术可被视为一种预防选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/349a/5875072/ec2486949568/ejifcc-18-091-g001.jpg

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