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Zhonghua Minguo wei sheng wu ji mian yi xue za zhi = Chinese journal of microbiology and immunology最新文献

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Autopsy findings on patients with AIDS in Taiwan. 台湾艾滋病患者的尸检结果。
C H Hsiao, S H Huang, S F Huang, C L Song, I J Su, C Y Chuang, Y T Yao, C T Lin, H C Hsu

At National Taiwan University Hospital, from 1986 to 1996, autopsies were performed on 16 patients with acquired immunodeficiency syndrome. There were 15 men and 1 woman. Fourteen of these male patients had contracted the disease as a result of sexual practice, among which 9 were homosexual, 1 was bisexual and 4 were heterosexual. One of the patients had become infected by sharing a syringe during intravenous drug use. The female was a sex worker. Among these patients, only 2 had been tested for HIV before developing AIDS. On autopsy, lymphoid depletion and thymus atrophy were found in all patients. Testicular atrophy was noted in all the male patients. Three patients died of malignant lymphoma. Twelve patients died of opportunistic infections and 1 committed suicide. The initial opportunistic infection was usually oral candidiasis. Pneumocystis carinii pneumonia (PCP) was the most common opportunistic infection developed in the early stage while cytomegalovirus (CMV) infection was the most common one found in the late stage. Mycobacterium infection had developed in 8 patients. Six patients had disseminated Kaposi's sarcoma (KS) and 4 of them were homosexual. In 4 patients, biopsy specimens were proved to have KS associated viral (HHV-8) genome. Malignant lymphoma was found in 4 cases, all were of high grade B cell type. Epstein-Barr virus (EBV) encoding small RNA (EBER1) was demonstrated in all the lymphomas. In conclusion, (1) the prevalence of tuberculosis (38%) in patients with AIDS in Taiwan is high; (2) the most common opportunistic infections in this series are candidiasis, PCP and CMV infections; (3) the incidence of AIDS related non-Hodgkin's lymphoma in Taiwan has increased since 1995.

在国立台湾大学医院,从1986年到1996年,对16名获得性免疫缺陷综合征患者进行了尸检。有15名男性和1名女性。其中14例男性患者因性行为感染该病,其中同性恋9例,双性恋1例,异性恋4例。其中一名患者因在静脉注射毒品时共用一个注射器而感染。这名女性是一名性工作者。在这些患者中,只有2人在患艾滋病之前接受过艾滋病毒检测。尸检发现所有患者均出现淋巴细胞减少和胸腺萎缩。所有男性患者均有睾丸萎缩。3例患者死于恶性淋巴瘤。12名患者死于机会性感染,1名患者自杀。最初的机会性感染通常为口腔念珠菌病。卡氏肺囊虫肺炎(PCP)是早期最常见的机会感染,而巨细胞病毒(CMV)感染是晚期最常见的机会感染。8例患者发生分枝杆菌感染。弥散性卡波西肉瘤(KS) 6例,其中4例为同性恋。在4例患者中,活检标本被证明具有KS相关病毒(HHV-8)基因组。恶性淋巴瘤4例,均为高分级B细胞型。所有淋巴瘤均可见编码小RNA (EBER1)的eb病毒(EBV)。综上所述:(1)台湾地区艾滋病患者结核病患病率较高(38%);(2)该系列中最常见的机会性感染是念珠菌病、PCP和CMV感染;(3)台湾地区AIDS相关非霍奇金淋巴瘤发病率自1995年以来呈上升趋势。
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引用次数: 0
Transfer of the E6 and E7 genes of human papillomavirus type 18 into human epithelial cells via recombinant retrovirus infection. 重组逆转录病毒感染人18型乳头瘤病毒E6和E7基因转染人上皮细胞
C J Wu, K S Chang, Y S Chang

A retroviral vector carrying the E6 and E7 genes of HPV type 18 was transfected into a packaging cell line, Ampho psi 2. Thirteen recombinant viruses carrying the E6 and E7 genes were obtained. The titers of these recombinant viruses were estimated by infecting BALB/c3T3 cells and then counting the number of G418r colonies. Presence of HPV E6/E7 genes was confirmed by the PCR method and sequence-specific primers. The expression of E7 gene was examined by RT-PCR method. Results showed that the titers were ranged between 0.2 and 1.2 x 10(3) CFU/ml and the E7 transcripts were detected in all 13 cell clones. These E6 and E7-containing cell clones were able to grow in soft agar, indicating the E6/E7 delivered by the recombinant retroviruses retained their transformation function. These recombinant viruses were then used to infect human NPC cell lines, NPC-TW076 and -TW039 and cell clones resistant to G418 were obtained. Using Western blot analysis and HPV type 18 E6-specific monoclonal antibody, HPV-CIP5, these cells were shown to contain a protein with a molecular mass of 18 kDa. Our data indicated that the HPV E6/E7-containing recombinant retroviruses were capable of infecting human cell lines. The potential of using these recombinant retroviruses to immortalize human primary epithelial cells was discussed.

将携带HPV 18型E6和E7基因的逆转录病毒载体转染到包装细胞系Ampho psi 2中。共获得13个携带E6和E7基因的重组病毒。通过感染BALB/c3T3细胞,计算G418r菌落的数量来估计重组病毒的滴度。通过PCR方法和序列特异性引物证实存在HPV E6/E7基因。采用RT-PCR法检测E7基因的表达。结果显示,E7的滴度在0.2 ~ 1.2 × 10(3) CFU/ml之间,13个克隆中均检测到E7转录本。这些含有E6和E7的细胞克隆能够在软琼脂中生长,表明重组逆转录病毒传递的E6/E7保留了它们的转化功能。利用重组病毒感染人鼻咽癌细胞株NPC- tw076和-TW039,获得了抗G418的细胞克隆。通过Western blot分析和HPV 18型e6特异性单克隆抗体HPV- cip5,发现这些细胞含有一个分子质量为18 kDa的蛋白。我们的数据表明,含有E6/ e7的重组逆转录病毒能够感染人类细胞系。讨论了利用这些重组逆转录病毒使人原代上皮细胞永生化的可能性。
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引用次数: 0
Strategies for diagnosing HIV-1 infection in atypical Western blots. 非典型免疫印迹法诊断HIV-1感染的策略。
M Y Chen, K L Lee, C C Hung, C Y Chuang, M J Chou

The Western blot (WB) has long been used to confirm positive ELISAs for diagnosing HIV-1 infections. However, some WB patterns may result in "indeterminate" or controversial reports thus impeding early diagnoses or accurate diagnoses. The interpretation of HIV-1 WB has no "gold standard" criterion. Incomplete antibody profiles on WB strips can be interpreted as positive or indeterminate according to different criteria. The possibility of HIV-2 infection was further checked in these serum samples. However, no reactivity to synthetic peptide of HIV-2 gp36 had been found. Serial WB analyses are important for attaining early diagnoses of HIV-1 infections as well as for evaluating clinical stages. Temporal changes on WB patterns of serial serum samples provide the evidence of seroconversion in individuals with risk behaviours and indeterminate WB. In late stage of HIV-1 infection, the reactivity to gag, pol and env antigen groups may decrease and result in indeterminate WB. We propose to diagnose HIV-1 infection and to differentiate the infection of HIV-1 from HIV-2 in these cases by using nested polymerase chain reaction (PCR) to demonstrate the presence of HIV-1 specific vpu gene.

长期以来,Western blot (WB)一直用于确认诊断HIV-1感染的阳性elisa。然而,一些WB模式可能导致“不确定”或有争议的报告,从而阻碍早期诊断或准确诊断。HIV-1 WB的解释没有“金标准”标准。根据不同的标准,WB条带上不完整的抗体谱可以解释为阳性或不确定。在这些血清样本中进一步检查HIV-2感染的可能性。但未发现对HIV-2 gp36合成肽具有反应性。连续WB分析对于获得HIV-1感染的早期诊断以及评估临床阶段非常重要。系列血清样本中血清白蛋白模式的时间变化为有危险行为和血清白蛋白不确定的个体提供了血清转化的证据。在HIV-1感染的晚期,对gag、pol和env抗原群的反应性可能降低,导致WB不确定。我们建议通过巢式聚合酶链反应(PCR)来证实HIV-1特异性vpu基因的存在,从而诊断HIV-1感染,并在这些病例中区分HIV-1与HIV-2感染。
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引用次数: 0
Arylamine N-acetyltransferase activity in Staphylococcus aureus. 金黄色葡萄球菌中芳胺n -乙酰转移酶活性。
F C Chang, J G Chung, W C Chang, L T Wu, G W Chen, S H Chang

N-Acetyltransferase (NAT) activities were determined by incubation of Staphylococcus aureus cytosols with p-aminobenzoic acid (PABA) or 2-aminofluorene (2-AF) followed by high pressure liquid chromatography assays. The NAT activities from S. aureus were found to be 0.67 +/- 0.04 nmol/min/mg protein for the acetylation of 2-AF and 0.46 +/- 0.02 nmol/min/mg protein for the acetylation of PABA. The apparent K(m) and Vmax values obtained were 2.85 +/- 0.65 mM and 7.51 +/- 0.86 nmol/min/mg protein for 2-AF, and 2.35 +/- 0.39 mM and 9.43 +/- 0.78 nmol/min/mg protein for PABA, respectively. The optimal pH value for the enzyme activity was 7.0 for both substrates tested. The optimal temperature for enzyme activity was 37 degrees C for both substrates. The NAT activity was inhibited by iodoacetamide at 0.25 mM, and activity was reduced 50%. At 1.0 mM iodoacetamide activity was inhibited more than 90%. Among a series of divalent cations and salts, Zn2+, Ca2+, and Fe2+ were demonstrated to be the most potent inhibitors. The molecular weight of NAT from S. aureus was found to be 44.9 kDa. This report is the first demonstration of acetyl CoA: arylamine NAT activity in S. aureus.

采用对氨基苯甲酸(PABA)或2-氨基芴(2-AF)孵育金黄色葡萄球菌细胞质,采用高压液相色谱法测定n -乙酰转移酶(NAT)活性。结果表明,金黄色葡萄球菌对2-AF乙酰化的NAT活性为0.67 +/- 0.04 nmol/min/mg,对PABA乙酰化的NAT活性为0.46 +/- 0.02 nmol/min/mg。2-AF的表观K(m)和Vmax分别为2.85 +/- 0.65 mM和7.51 +/- 0.86 nmol/min/mg蛋白,PABA的表观K(m)和Vmax分别为2.35 +/- 0.39 mM和9.43 +/- 0.78 nmol/min/mg蛋白。两种底物酶活性的最佳pH值均为7.0。两种底物酶活性的最佳温度均为37℃。0.25 mM碘乙酰胺对NAT活性有抑制作用,活性降低50%。1.0 mM时,碘乙酰胺活性被抑制90%以上。在一系列二价阳离子和盐中,Zn2+、Ca2+和Fe2+被证明是最有效的抑制剂。从金黄色葡萄球菌中分离得到的NAT分子量为44.9 kDa。本文首次证实了乙酰辅酶a:芳胺在金黄色葡萄球菌中的NAT活性。
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引用次数: 0
Production and purification of Bordetella pertussis toxin. 百日咳杆菌毒素的制备与纯化。
C L Ju, G C Sheu, Y Cheng, C H Lu

Pertussis toxin (PT) is the major protective antigen of acellular pertussis vaccine (aP). We have established an optimal culture condition for the growth of B. pertussis and the production of PT in a laboratory scale fermentor. It was found that when the dissolved oxygen in medium was supplied with pure oxygen instead of air, the yield of PT was dramatically increased (i.e. from 2-3 mg/l using air to 8-10 mg/l using pure oxygen). PT was purified by affinity chromatography using hydroxyapatite and fetuin-sepharose columns. SDS-PAGE analysis and CHO cell clustering test showed that the purified PT was comparable to the reference PT in purity and biological activity. The purified PT could be detoxified by formaldehyde (d-PT). The results of CHO cell clustering neutralization assay and ELISA showed that the antibody induced by d-PT in mice was comparable to that induced by PT contained in a commercial DTaP. These results indicated that the immunogenicity of our d-PT was retained after the purification and detoxification procedures.

百日咳毒素(PT)是无细胞百日咳疫苗(aP)的主要保护性抗原。在实验室规模的发酵罐中建立了百日咳双球菌生长和生产PT的最佳培养条件。实验发现,当介质中的溶解氧以纯氧代替空气供给时,PT的产率显著提高(即由空气供给的2 ~ 3 mg/l提高到纯氧供给的8 ~ 10 mg/l)。PT采用羟基磷灰石和葡糖精亲和层析纯化。SDS-PAGE分析和CHO细胞聚类试验表明,纯化后的PT在纯度和生物活性方面与参考PT相当。纯化后的PT可以用甲醛(d-PT)解毒。CHO细胞聚类中和实验和酶联免疫吸附试验结果表明,d-PT在小鼠体内诱导的抗体与商业DTaP中含有的PT诱导的抗体相当。这些结果表明,我们的d-PT的免疫原性在纯化和解毒程序后保持不变。
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引用次数: 0
Prevalence and heterogeneity of Helicobacter pylori in gastric biopsies of patients with gastroduodenal diseases. 胃十二指肠疾病患者胃活检中幽门螺杆菌的患病率和异质性。
C W Lin, Y S Chang, P Y Lai, K S Cheng

Helicobacter pylori is a risk factor for development of peptic ulcers and adenocarcinoma of distal stomach. There are several highly specialized virulence factors, such as the production of sialic acid-specific hemagglutinins, cytotoxins and enzymes. This study was designed to study the in vivo prevalence of H. pylori in patients with gastroduodenal diseases, the in vivo correlation between H. pylori infection and blood group O, and the heterogeneity of H. pylori isolates in central Taiwan. We enrolled 776 symptomatic patients residing in the central Taiwan area. The age-specific in vivo prevalence of H. pylori in patients with gastroduodenal diseases increased from 11.1% in those between the ages of 1 to 20, 73.1% in those between the ages of 21 and 30, and to 79.8% in those between the ages of 51 and 60. In conclusion, H. pylori was present in 70% of biopsied specimens of symptomatic patients with gastroduodenal diseases and had the highest incidence (86%) in patients with peptic ulcers. The prevalence of H. pylori cag A expression positive strains in central Taiwan was 92.5%. This study has also demonstrated the high correlation between H. pylori and the blood group O-positive patients with gastroduodenal diseases. The prevalence of H. pylori infection in blood O-positive patients in central Taiwan was 86.4%.

幽门螺杆菌是消化性溃疡和远端胃腺癌发展的危险因素。有几个高度专业化的毒力因素,如唾液酸特异性血凝素,细胞毒素和酶的生产。本研究旨在探讨台湾中部地区胃十二指肠疾病患者体内幽门螺杆菌的流行情况、体内幽门螺杆菌感染与O型血的相关性以及幽门螺杆菌分离株的异质性。我们招募了居住在台湾中部地区的776名有症状的患者。胃十二指肠疾病患者体内年龄特异性幽门螺杆菌患病率从1 ~ 20岁的11.1%、21 ~ 30岁的73.1%、51 ~ 60岁的79.8%上升。总之,有症状的胃十二指肠疾病患者的活检标本中有70%存在幽门螺杆菌,其中消化性溃疡患者的发生率最高(86%)。台湾中部地区幽门螺杆菌cag A阳性表达株的检出率为92.5%。本研究也证实了幽门螺杆菌与o型血阳性胃十二指肠疾病患者之间的高度相关性。中部地区血o型阳性患者幽门螺杆菌感染率为86.4%。
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引用次数: 0
Clinical experience of HIV/AIDS in a municipal hospital in Taiwan. 台湾某市立医院HIV/AIDS的临床经验。
Y L Wu, C C Chang, H C Chang, C H Yang, S T Liao

To describe the spectrum of epidemiological and major clinical manifestations of patients infected by human immunodeficiency virus type 1 (HIV-1) in a municipal hospital, a retrospective review was done of 53 HIV-1-infected patients who had been admitted to Taipei Municipal Jen-Ai Hospital between January 1990, and July 1996. The majority (94.3%) of the patients in the cohort were male. Peak incidence was found in the fourth decade (28.3%). Forty-four (83%) patients presented in the first hospital stay with acquired immunodeficiency syndrome (AIDS). The mean duration between establishment of diagnosis of HIV-1 infection and that of AIDS was 11.2 (0-84) months. Heterosexual transmission accounted for 54.7% of the infections in the study group, and bi-/homosexual men made up another 32%. Psychosis of new onset was noted in two patients. In all AIDS indicator conditions, Pneumocystis carinii pneumonia (PCP) was the leading opportunistic infection among AIDS patients. PCP was also on the top of initial manifestations of HIV-1 infection. One patient with Penicillium marneffei infection was diagnosed to have AIDS. The mean CD4 count at admission of AIDS patients was much lower than that of non-AIDS patients (32 vs. 297/microliter, p < 0.0005). During the follow-up period 24 of 53 patients died. Mean survival time of 23 expired patients after establishment of diagnosis of AIDS was 6.4 (0-29) months. The results indicated that males outnumbered females greatly in the number of cases. Sexual activity remained the most important route of infection. Psychosis of new onset may be an early manifestation of HIV-associated encephalopathy and requires more attention. In addition, the outcome was poor as most patients in this area did not become aware of risk of HIV-1 infection until they were seriously illed with full-blown AIDS that they would seek medical help. PCP was the most common incentive for medical consultation. Penicillium marneffei infection is endemic in southeast Asia, and should be classified as an AIDS indicator condition in Taiwan.

摘要为了描述市立医院感染人类免疫缺陷病毒1型(HIV-1)患者的流行病学特征和主要临床表现,我们对1990年1月至1996年7月间在台北市仁爱医院收治的53例HIV-1感染患者进行了回顾性分析。该队列中大多数患者(94.3%)为男性。发病率高峰出现在第四个十年(28.3%)。44例(83%)患者首次住院时出现获得性免疫缺陷综合征(AIDS)。从诊断为HIV-1感染到诊断为艾滋病的平均时间为11.2(0 ~ 84)个月。在研究组中,异性恋传播占感染的54.7%,双性恋/同性恋男性占32%。2例患者出现新发精神病。在所有艾滋病指标条件下,卡氏肺囊虫肺炎(PCP)是艾滋病患者中主要的机会性感染。PCP在HIV-1感染的初始表现中也居首位。一名感染马尔尼菲青霉的患者被诊断为艾滋病。艾滋病患者入院时平均CD4计数明显低于非艾滋病患者(32 vs 297/微升,p < 0.0005)。在随访期间,53例患者中有24例死亡。23例确诊后死亡患者的平均生存时间为6.4(0 ~ 29)个月。结果表明,男性病例数大大超过女性。性行为仍然是最重要的感染途径。新发精神病可能是hiv相关脑病的早期表现,需要更多的关注。此外,结果也很差,因为该地区的大多数患者直到艾滋病严重发展到需要寻求医疗帮助时才意识到感染艾滋病毒-1的风险。PCP是最常见的医疗咨询动机。马尔尼菲青霉感染是东南亚地区的一种地方病,在台湾应被列为艾滋病的指示性疾病。
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引用次数: 0
Molecular subtyping of the HIV-1 V3 loop sequences detected in HIV-1-positive patients in southern Taiwan. 台湾南部HIV-1阳性患者HIV-1 V3环序列的分子分型。
B Shieh, C M Lee, Y M Chen, I J Su, C Li

Polymerase chain reaction and nucleotide sequence analysis were performed to amplify and determine the V3 loop sequences of human immunodeficiency virus type 1 (HIV-1) from ten seropositive patients at National Cheng Kung University Hospital, Tainan. The nucleotide sequences and the deduced amino acid (a. a.) sequences of these V3 regions were compared with those of known HIV-1 prototypes. The V3 loop a. a. sequences detected in eight individuals belong to subtype B which predominates in North America and Europe, whereas two individuals were infected with HIV-1 subtype E which is mainly found in the heterosexual populations of Thailand. Sequence analysis of these variant HIV-1 strains revealed a number of interesting features and a phylogenetic tree was also constructed according to the V3 loop nucleotide sequences of these variant strains and HIV-1 isolates from other parts of the world. Furthermore, our results suggest that the north vs south geographical separation in terms of HIV-1 epidemiology in Taiwan is insignificant.

采用聚合酶链反应和核苷酸序列分析方法,对台南国立成功大学医院10例血清阳性患者的HIV-1型病毒V3环序列进行扩增和测定。将这些V3区域的核苷酸序列和推导出的氨基酸序列与已知的HIV-1原型进行比较。在8个个体中检测到的V3环a. a.序列属于在北美和欧洲占主导地位的B亚型,而2个个体感染了主要在泰国异性恋人群中发现的HIV-1 E亚型。对这些变异HIV-1毒株的序列分析揭示了许多有趣的特征,并根据这些变异毒株和世界其他地区HIV-1分离株的V3环核苷酸序列构建了系统发育树。此外,本研究结果显示台湾HIV-1流行病学的南北地理差异不显著。
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引用次数: 0
[Primary surveillance of spotted fever group antibodies on rats in the Kinmen area]. 金门地区大鼠斑点热群抗体初步监测。
H L Chen, H Y Chen, C L Chung, T H Lin, G R Wang, C B Horng

The positive rate of rickettsial antibodies of 107 rats in the Kinmen area by indirect immunofluorescent antibody (IFA) technique was 0% (0/107) in typhus fever, 38.3% (41/107) in scrub typhus and 66.4% (71/107) in spotted fever group; the positive rate (42.9%) of spotted fever group of 21 rats in Taiwan island also higher than scrub typhus (19.0). It suggests that spotted fever group patients may be present in our country but have not been discovered.

间接免疫荧光抗体(IFA)法检测金门地区107只大鼠的立克次体抗体阳性率分别为斑疹热组0%(0/107)、恙虫病组38.3%(41/107)和斑疹热组66.4% (71/107);台湾岛斑点热组21只大鼠阳性率(42.9%)高于恙虫病(19.0%)。提示我国可能存在斑点热组患者,但尚未发现。
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引用次数: 0
Immunogenicity of Haemophilus influenzae b conjugate vaccine (HibTITER) and safety of HibTITER and a combination vaccine of diphtheria, tetanus, pertussis and HibTITER in infants two months of age: a preliminary report. b型流感嗜血杆菌结合疫苗(hititer)的免疫原性以及hititer和白喉、破伤风、百日咳和hititer联合疫苗在两个月龄婴儿中的安全性:初步报告
P F Chang, L M Huang, P I Lee, H H Chiu, S Y Tasi, C Y Lee

A study was undertaken to evaluate the safety and immunongenicity of a conjugate Haemophilus influenzae type b (Hib) vaccine (HibTITER) when administered concurrently with DTP (diphteria, tetanus and pertussis) vaccine in separate syringes. A total of 90 healthy children (45 per group) were randomized to receive either TETRAMUNE, a vaccine combining HibTITER with whole-cell DTP (group A), or DTP and HibTITER administered concurrently (group B) in separate syringes at approximately 2, 4 and 6 months of age in Taiwan. All children in group B achieved anti-Hib PRP (polyribosylribitol phosphate) antibody titers above 0.15 microgram/ml and 91% developed antibody titers above 1.0 microgram/ml following the third immunization. Incidences of adverse reactions were comparable between groups A and B. Besides, the incidences of adverse reactions were not significantly more frequent compared with DTP vaccination alone. We concluded that HibTITER was highly immunogenic and safe when administered concurrently with DTP vaccine to Taiwanese children. TETRAMUNE was also safe and the number of injections may be reduced in the future.

进行了一项研究,以评估b型流感嗜血杆菌(Hib)结合疫苗(hititer)与百白破(白喉、破伤风和百日咳)疫苗在单独注射器中同时使用时的安全性和免疫原性。在台湾,共有90名健康儿童(每组45名)随机接受TETRAMUNE,一种将HibTITER与全细胞百白破联合接种的疫苗(A组),或在大约2、4和6个月大时,在单独的注射器中同时接种DTP和HibTITER (B组)。B组儿童抗hib PRP抗体滴度均高于0.15微克/毫升,91%的儿童在第三次免疫后抗体滴度高于1.0微克/毫升。不良反应发生率在A组和b组之间具有可比性,而且不良反应发生率与单独接种百白破疫苗相比没有明显增加。我们的结论是,当与百白破疫苗同时使用时,HibTITER对台湾儿童具有高度的免疫原性和安全性。TETRAMUNE也是安全的,未来可能会减少注射次数。
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引用次数: 0
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Zhonghua Minguo wei sheng wu ji mian yi xue za zhi = Chinese journal of microbiology and immunology
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