新生儿肺炎死亡病例中B族链球菌的检测

目的对北京儿童医院新生儿肺炎死亡病例进行B族链球菌（GBS）回顾性检测,旨在初步揭示GBS感染在新生儿重症肺炎中的地位。方法收集200例1953年到2004年间新生儿肺炎死亡病例（研究组）和34例非感染性疾病新生儿死亡病例（对照组）的尸检病理标本,提取石蜡组织DNA,通过PCR和Southem blot技术检测GBS特异性基因片断咖基因。同时对所选病例的临床资料进行复习。结果（1）研究组200例中,PCR对GBS检出率为26％,Southern blot检出率为65％,检出率均明显高于对照组（PCR3％,x^2=8．82,P〈0．01;Southern blot,18％,x^2=26．77,P〈0．01）。（2）研究组中,年龄小于7d的病例（早发型）103例,年龄大于7d的病例（晚发型）97例,小于7d的患儿GBS检出率明显高于大于7d的患儿,其中PCR检出率在前者为37％,后者为13％（x^2=15．537,P〈0．01）,Southern blot检出率在前者为72％,后者为52％（x^2=4．37,P〈0．05）。早发型阳性病例中,39％为早产儿（29／74）。（3）研究组中,75例有完整临床资料,其中35例可查到一项或多项与GBS感染有关的危险因素,如早产,低体重,胎膜早破,羊水异常等,GBS检测结果均为阳性。阳性病例中,早发型最常见的表现有青紫、窒息,呼吸困难,晚发型最常见的表现有咳喘,呼吸困难。对照组中,1例PCR检测阳性者患恶性畸胎瘤,其他5例仅Southern blot检测阳性者分别为核黄疸、肝癌、先天性无肛合并膀胱尿道瘘、新生儿自然出血、先天性低位无肛合并直肠会阴瘘患者。结论GBS是新生儿肺炎死亡病例中的重要病原,尤其在早发型肺炎病例中GBS感染占很大比重。Southern blot是在石蜡标本中检测GBS的较为敏感方法。

Detection of group B streptococcus in the cases died of neonatal pneumonia

OBJECTIVE: From the 1970s, group B streptococci (GBS) have been widely recognized as an important pathogen in neonatal infectious disease, and it emerged as the leading cause of neonatal morbidity and mortality in the Western world. However, there are few data on the prevalence of neonatal GBS infections in China. The aim of this retrospective study was to estimate whether GBS is an important pathogen in severe neonatal pneumonia, and to develop a method for detection of GBS infections in fatal neonatal pneumonia. METHODS: A total of 234 neonatal cases (0 - 28 days) died in Beijing Children's Hospital from 1953 to 2004 were enrolled in this study. They were divided into two groups. Two hundred cases diagnosed as neonatal pneumonia were assigned to study group and the remaining 34 cases died of neonatal hemolysis or surgical operation without any confirmed infectious diseases were designated as control group. Formalin-fixed, paraffin-embedded lung tissues were used as source for total genomic DNA extraction. PCR and Southern blot analyses were applied to detect GBS specific cfb gene target sequence. And the clinical data of these cases were reviewed as well. RESULTS: In the study group, 52 cases were detected positive for GBS DNA by PCR (26%), 130 cases were positive by Southern blot (65%). In the control group, 1 case was detected positive GBS DNA by PCR (3%), and 6 cases were positive by Southern blot (18%). The positive rate was significantly lower in the control group than that in the study group (PCR, chi(2) = 8.82, P < 0.01; Southern blot, chi(2) = 26.77, P < 0.01). The positive rate in the neonates younger than 7 days (early-onset) was significantly higher than that in neonates older than 7 days (late-onset) (PCR: 37% vs. 13%, chi(2) = 15.537, P < 0.01; Southern blot: 72% vs. 52%, chi(2) = 4.37, P < 0.05). In the positive early-onset cases, 39% of whom were born prematurely (29/74). Out of the 200 cases, 75 had complete clinical data. Neither blood nor lung culture for GBS was performed in any of these cases. But risk factors were identified for 35 cases, such as premature delivery, low birth weight, premature rupture of the membrane and abnormal amniotic fluid. GBS was positive in all these cases. Severe apnea appeared to be a common symptom and was present in most of the early-onset GBS-positive cases, while cough and wheezing were found in most of the late-onset GBS-positive cases. In the control group, one PCR positive case was suffered from malignant teratoma. The other 5 positive cases confirmed by Southern blot were diagnosed as kernicterus, hepatoma, aproctia complicating with cysti-urethral fistula, neonatal physio logical bleeding and aproctia complicated with archo-perineal fistula. CONCLUSION: Group B Streptococcus is an important pathogen in fatal neonatal pneumonia, especially in early-onset cases. southern blot may be a sensitive method to detect GBS infection in archival tissues. In the clinical work, more attention should be paid to the neonates with GBS risk factors. And GBS detection and prevention in neonates should be put into clinical practice.