卡氏肺孢子虫感染的免疫诊断研究

目的：探讨肺泡灌洗液、血清抗原及血清抗体检测诊断卡氏肺孢子虫感染的价值。方法：利用免疫抑制大鼠模型及双夹心ＥＬＩＳＡ法检测不同时期感染大鼠肺泡灌洗液和血清中肺孢子虫抗原，用ＩＦＡ法检测血清中的肺孢子虫ＩｇＧ抗体，并与病原学检查结果进行比较。结果：感染大鼠肺泡灌洗液的抗原检测于免疫抑制６－８ｗｋ后均呈阳性，而对照组大鼠均呈阴性；大多数感染大鼠血清抗原检测为阴性；正常大鼠血清中有低滴度肺孢子虫ＩｇＧ抗体，感染大鼠抗体滴度轻度升高或不升高，但中止免疫抑制后血清抗体滴度明显升高，而肺泡灌洗液中抗原逐渐阴转。结论：肺泡灌洗液抗原检测可用于肺孢子虫感染的诊断，但血清中很难检出这种抗原；血清ＩｇＧ抗体的上升并不表示为现症感染。     

SPA-ELISA检测卡氏肺孢子虫抗原的研究

目的探讨SPAK-ELISA检测卡氏肺孢子虫抗原对诊断卡氏肺孢子虫肺炎的价值。方法采用SPAELISA检测免疫抑制大鼠模型在不同时期肺泡灌洗液(BALF)和血清中肺孢子虫(PC)抗原，以及造血干细胞移植并发肺炎患者痰液和血清抗原，并与病原学检查结果比较。结果感染大鼠于用药后第4周，BALF中PC抗原，以及肺组织印片和BALF涂片包囊检查开始出现阳性，但前者阳性率(60％)高于后者(20％和40％)，而且全部阳性达到的时间(8周)也早于后者(10周)。第10周有2只鼠血清抗原阳性，但此后又转阴。30例肺炎患者痰液抗原阳性8例，阳性率为26．67％，8例抗原阳性患者中只有3例发现包囊，符合率为37．5％。血清抗原检测全部阴性。结论SPA—ELISA检测痰液和肺泡灌洗液抗原比病原学检查敏感性高，而检测血清抗原不适用于诊断卡氏肺孢子虫肺炎。     

SPA-ELISA检测卡氏肺孢子虫抗原的研究

目的 探讨SPAK-ELISA检测卡氏肺孢子虫抗原对诊断卡氏肺孢子虫肺炎的价值。 方法 采用SPA-ELISA检测免疫抑制大鼠模型在不同时期肺泡灌洗液(BALF)和血清中肺孢子虫(PC)抗原,以及造血干细胞移植并发肺炎患者痰液和血清抗原,并与病原学检查结果比较。 结果 感染大鼠于用药后第4周,BALF中PC抗原,以及肺组织印片和BALF涂片包囊检查开始出现阳性,但前者阳性率(60％)高于后者(20％和40％),而且全部阳性达到的时间(8周)也早于后者(10周)。第10周有2只鼠血清抗原阳性,但此后又转阴。30例肺炎患者痰液抗原阳性8例,阳性率为26.67％,8例抗原阳性患者中只有3例发现包囊,符合率为37.5％。血清抗原检测全部阴性。 结论 SPA-ELISA检测痰液和肺泡灌洗液抗原比病原学检查敏感性高,而检测血清抗原不适用于诊断卡氏肺孢子虫肺炎。     

大鼠实验性卡氏肺孢子虫肺炎中IL-2,IL-6的变化

目的 建立大鼠实验性卡氏肺孢子虫肺炎的模型,观察血清和肺泡灌洗液中IL-2和IL-6的变化.方法 给SD大鼠皮下注射地塞米松磷酸钠建立卡氏肺孢子虫肺炎动物模型,酶免法检测支气管肺泡灌洗液中IL-2和IL-6含量及血清中IL-6含量;放免法检测血清中IL-2含量.结果 感染组大鼠血清和肺泡灌洗液中IL-2和IL-6含量均显著高于正常对照组.结论 卡氏肺孢子虫肺炎大鼠血清和肺泡灌洗液中IL-2和IL-6升高.     

蒿甲醚用于卡氏肺孢子虫肺炎大鼠的治疗及其对IL-6影响的研究

目的 从病理学和细胞因子角度探讨蒿甲醚对大鼠卡氏肺孢子虫肺炎（Pneumocystiscarinii Pneumonia POP）的治疗效果和作用机理。方法 给SD大鼠皮下注射地塞米松磷酸钠建立卡氏肺孢子虫肺炎动物模型，治疗组给予蒿甲醚治疗。ELISA双抗夹心法分别检测血清和支气管肺泡灌洗液中1L-6水平。结果 与感染对照组比较，蒿甲醚治疗组症状显著改善、肺印片中卡氏肺孢子虫包囊数目显著减少、肺组织炎症明显减轻、血清和肺泡灌洗液中IL-6水平明显下降。结论 蒿甲醚具有一定抗大鼠卡氏肺孢子虫肺炎作用，能够降低PCP大鼠IL-6。     

血清曲霉IgG抗体联合支气管肺泡灌洗液半乳甘露聚糖检测对肺曲霉菌病诊断价值分析

目的探讨血清曲霉IgG抗体联合肺泡灌洗液(BALF)半乳甘露聚糖(GM)检测对肺曲霉菌感染的诊断价值,为肺曲霉菌病的临床诊断提供新的方案。方法选择2016年3月至2017年9月首都医科大学附属北京朝阳医院收治的疑似肺曲霉菌感染患者97例,对患者进行侵袭性肺曲霉菌病(IPA)和慢性肺曲霉菌病(CPA)诊断,同时检测患者血清曲霉IgG抗体以及BALF GM抗原,采用ROC曲线分析各检测方法对肺曲霉菌病的诊断价值。结果97例患者中,肺曲霉菌病患者确诊9例(9.28%)、临床诊断49例(50.52%)、排除肺曲霉菌感染患者39例(40.21%),将确定诊断、临床诊断患者58例作为病例组,排除肺曲霉菌感染患者39例作为对照组。病例组患者血清IgG抗体及BALF GM抗原水平均显著高于对照组(P0.05),其中CPA患者血清IgG抗体显著高于IPA患者(P0.05)。血清曲霉IgG以140 kAU/L为临界值时,敏感度为53.4%,特异度为94.9%。支气管肺泡灌洗液GM以0.5为临界值时,敏感度为75.9%,特异度为76.9%。单独采用血清曲霉IgG抗体对肺曲霉菌病诊断曲线下面积为0.770,单独采用BALF GM抗原对肺曲霉菌病诊断曲线下面积为0.813,二者联合对肺曲霉菌病诊断曲线下面积为0.897。结论血清曲霉IgG抗体联合血清半乳甘露聚糖检测可有效提高对肺曲霉菌病患者的临床诊断价值,有助于患者的临床诊断,值得推广运用。     

用漱口液PCR法无创伤性诊断卡氏肺孢子虫性肺炎

除了艾滋病患者外,血液病或骨髓移植患者也好发卡氏肺孢子虫性肺炎(PCP)。PCP的诊断通常需要患者支气管肺泡灌洗 液或吸痰液,而支气管肺泡的灌洗比较困难,吸痰液的诊断敏感性差别较大。 对HIV-1阳性合并卡氏肺孢子虫性肺炎的患者,曾用标准PCR法检测到漱口液中的卡氏肺孢子虫的DNA,敏感性为78％。本文作者检测了26名血液病和有呼吸道症状     

DNA扩增法检测卡氏肺孢子虫

以寡核苷酸引物和探针用于多聚酶链反应(PCR)中,用来检测卡氏肺孢子虫特异的DNA片段。共检测47份诊断性支气管肺泡灌洗液标本,其中37份来自因人类免疫缺陷病毒(HIV)感染或免疫抑制剂治疗而处于免疫抑制状态者,全部患者都有急性呼吸道症     

肺泡灌洗液联合mNGS在免疫抑制合并肺部感染病原学的应用

目的 了解肺泡灌洗液联合宏基因组下一代测序技术(mNGS)在免疫抑制合并肺部感染中病原学诊断的应用价值。方法 回顾分析2020年1月-2022年5月确诊为肺部感染的免疫抑制患者71例及非免疫抑制患者167例的临床资料，同时收集患者痰培养/痰涂片、肺泡灌洗液联合mNGS病原学的检出结果，针对肺泡灌洗液联合mNGS在免疫抑制合并肺部感染患者中的诊断及治疗指导效能进行分析，并绘制ROC曲线。结果 免疫抑制组中肺泡灌洗液联合mNGS检出病原体阳性率高于常规痰培养，差异有统计学意义(P<0.05); CRP、PCT、住院费用、住院天数等指标免疫抑制组均高于非免疫抑制组，淋巴细胞绝对计数低于非免疫抑制组(P<0.001)。将免疫抑制组分为4个亚组，其中肾移植组住院天数及住院费用均高于外在激素类免疫抑制组(P<0.05);而淋巴细胞绝对计数及中位年龄均低于外在激素类免疫抑制组(P<0.05);免疫抑制组各亚组间肺泡灌洗液联合mNGS、痰培养与预后之间关系无统计学差异(P>0.05)。肺泡灌洗液联合mNGS阳性患者的预后优于阴性的患者，差异有统计学意义(P<0.00...     

用PCR扩增技术检测卡氏肺孢子虫DNA

本文报道了用ＰＣＲ扩增技术检测卡氏肺孢子虫感染大鼠的肺组织，支气管灌洗液和外周血病原体ＤＮＡ，并和肺印片结果进行比较。共检测了实验感染鼠３４只及正常大鼠６只，肺印片阳性者１８例，ＰＣＲ检测肺组织样品出现特异的扩增条带者１９例，检测支气管灌洗液样品出现特异的扩增条带者１６例，外周血样品全部阴性。根据实验结果，认为引法用于检测卡氏肺孢子虫病原体ＤＮＡ有很好的应用前景。     

Course of antibody production by the DIG-ELISA method in neonatally infected and juvenile infected rats after primary infection with Breinlia booliati (Filarioidea: Onchocercidae).

The course of antibody production in Wistar neonatal and juvenile rats after primary infection with Breinlia booliati was studied by the DIG-ELISA technique using filter papers impregnated with capillary blood drawn from the infected rat tails at 7, 14, 28, 60 and 90 days post infection. Sera of neonatally infected rats did not react with adult worm antigen until day 7 and the titers of antibody remained at very low levels for the next 7 days. There was little tendency to eliminate the filarial larvae during this time. The antibody levels then rose rapidly throughout the next fortnight and increased to a maximum at day 60 after which the titer leveled out at a constant high value until early patency at day 90. On the other hand, antibodies could be detected in sera of juvenile infected rats as early as day 7 and the levels of antibody rose markedly to a maximum at day 28. During the period from day 60 to day 90 at early patency, the antibodies declined gradually to lower levels. The humoral immune responses of 42 neonatally infected rats and 53 juvenile infected rats of 3 strains (Lewis, Wistar and Sprague Dawley) were tested against soluble B. booliati antigens from both female (1:50) and male (1:10) worm extracts by the DIG-ELISA method. Antibodies were detected in sera from all the microfilaremic and amicrofilaremic rats belonging to neonatally and juvenile infected groups. Sera of clean neonatal rats did not give a positive reaction zone.     

Serological cross-reaction among Bartonella henselae, Chlamydia pneumoniae and Coxiella burnetii by indirect fluorescence antibody method]

We studied the serological cross-reactions among Bartonella henselae, Chlamydia pneumoniae and Coxiella burnetii by indirect fluorescence antibody (IFA) method, using sera from 8 patients with cat scratch disease (CSD), 13 patients with C. pneumoniae infection and 12 patients with acute Q fever. B. henselae IgG antibody was negative in 13 patients with C. pneumoniae infection, and was positive in 3 (titers being 1:64) of 12 patients with Q fever, whereas B. henselae IgM antibody was negative in all the patients with C. pneumoniae infection or Q fever. C. burnetii IgG antibody was removed by absorption of these 3 sera with C. burnetii antigens, whereas B. henselae IgG antibody did not change. C. pneumoniae IgG antibody was positive in 3 (titers being 1:125 in two, 1:32 in one) of 8 patients with CSD. Both C. pneumoniae and B. henselae IgG antibody titers were significantly reduced by absorption of these 3 sera with B. henselae antigens. C. burnetii IgG or IgM antibodies were negative in all patients with CSD. In conclusion, no serological cross-reaction between B. henselae and C. burnetii was observed. On the other hand. B. henselae IgG antibody cross-reacted to C. pneumoniae antigens, whereas C. pneumoniae IgG antibody did not cross-react to B. henselae antigens. Our findings suggest that determination of B. henselae IgG or IgM antibodies were not influenced by C. pneumoniae and C. burnetii antigens.     

Immunological relationships of Long Island isolates of Babesia microti.

Studies to detect strain differences among two rodent-derived and one human-derived Babesia microti isolates from Long Island were undertaken, using various methods. Superinfection experiments using the homologous and heterologous isolates showed cross-protection. All hamsters were resistant to superinfection challenges of increasing dosages of both the homologous and heterologous isolates. Attempts to infect other laboratory animals with the Long Island isolates of B. microti were successful in intact and splenectomized Sprague-Dawley rats and questionable in Swiss mice. Nylar and CFW mice as well as CFW and Wistar intact and splenectomized rats were refractory to B. microti isolates from Long Island. Indirect fluorescence tests using convalescent sera from six Long Island cases of babesiosis showed no titer differences with tests using the three Long Island antigens as well as the Gray strain antigens. The rise of hamster IgG anti-B. microti antibody was followed by indirect immunofluorescence done at different parasitemia levels. The IgG antibody in hamsters was detected early in the course of infection, rose rapidly concurrent with increasing parasitemia, and became stable at high titers for the duration of the infection. IgG antibody titers were unaffected by homologous superinfection challenges.     

Serum IgG and IgA antibody responses to Campylobacter pylori in a group of healthy asymptomatic volunteers

Sera from 17 healthy asymptomatic volunteers were tested for presence of IgG and IgA antibodies against Campylobacter pylori and correlated with endoscopic biopsy findings. Three volunteers infected with C. pylori had the highest IgG antibody titers of the group. None of 14 C. pylori free subjects had significant IgG antibody levels. IgA antibody titers were negative in all subjects regardless of state of infection, in contrast to control sera from symptomatic C. pylori infected patients who manifested high IgA antibody levels.     

Cell-mediated and humoral immune responses in capuchin monkeys infected with Schistosoma japonicum or Schistosoma mansoni

Cell-mediated immune responses, assessed by lymphocyte clonal expansion in vitro, as well as humoral responses, assessed by an enzyme-linked immunosorbent assay (ELISA), were evaluated in capuchin monkeys during a 7-month infection with Schistosoma mansoni or with a Japanese or Philippine strain of Schistosoma japonicum. Although mounting a vigorous antibody response against parasite antigens, the S. mansoni-infected monkeys failed to show lymphocyte proliferation in response to stimulation with soluble adult worm antigen or soluble egg antigen derived from S. mansoni. Monkeys infected with S. japonicum responded to parasite antigens obtained from S. japonicum both by antibody production and lymphocyte blastogenesis. Monkeys infected with S. japonicum (Japanese strain) never developed detectable levels of circulating immune complexes (CIC). On the other hand high levels of CIC appeared at 7 months of infection in the monkeys infected with S. mansoni. The CIC levels exhibited negative correlations with intensity of infection. In studies of antigen species specificity, sera from S. mansoni-infected monkeys showed much higher IgG antibody titers to antigens derived from S. mansoni than to S. japonicum-derived antigens. On the other hand, monkeys infected with S. japonicum had comparable IgG antibody titers to antigens of both schistosome species.     

Seroepidemiologic evaluation of anti-toxic and anti-colonization factor immunity against infections by LT-producing Escherichia coli in rural Bangladesh

To evaluate serologic immunity against clinical infections by heat-labile enterotoxin-producing Escherichia coli (LT-ETEC) in rural Bangladesh, 124 children and adult women with LT-ETEC diarrhea (cases) were compared with 347 age-matched community controls. In paired acute-convalescent sera from the cases, IgG anti-CFA I and anti-CFA II antibody titers increased eight-to ninefold after infection by LT-ETEC with the homologous CFA, and IgG anti-LT antibody titers increased fourfold for all LT-ETEC infections. Anti-CFA and anti-LT titers peaked in controls aged 12-23 months, the age group with the highest incidence of ETEC infections. However, antibody titers were similar in acute sera from cases and in sera from controls. Although serum IgG anti-CFA and anti-LT antibodies rose in response to LT-ETEC infections and paralleled the age-specific incidence of ETEC in the community, these antibodies were not associated with a lower risk of LT-ETEC diarrhea.     

建立和评价ELISA法检测血清和唾液中旋毛虫IgG抗体的研究

目的 建立检测血清和唾液中旋毛虫抗体的酶联免疫吸附试验(ELISA)方法.方法 应用方阵滴定法,筛选适宜的旋毛虫抗原(肌肉幼虫可溶性抗原、肌肉幼虫排泄分泌抗原、成虫可溶性抗原、成虫排泄分泌抗原)浓度、血清和唾液稀释度、辣根过氧化物酶标记的山羊抗兔、山羊抗人IgG抗体稀释度.共有20份旋毛虫感染兔、10份旋毛虫病患者血清和唾液用于这4种抗原的敏感性试验,20份健康兔与38份其他寄生虫感染兔和患者的血清和唾液用于这4种抗原的特异性试验.结果 这4种抗原适宜包被浓度依次为8.0 μg/ml、6.0 μg/ml、10.0 μg/ml、9.0 μg/ml.适宜的血清稀释度依次为1:100、1:200、1:50、1:200,唾液均用原液.适宜的山羊抗兔、山羊抗人IgG稀释度分别为1:2 500和l:2 000.这4种抗原检测旋毛虫感染兔血清和唾液的敏感性分别为100%和80%~100%,检测旋毛虫病患者血清和唾液的敏感性分别为100%和60%~80%;检测血清和唾液的特异性依次为81.03%、89.65%、77.59%、82.76%和93.10%、96.55%、89.65%、91.35%.结论 建立了检测兔和人血清及唾液中旋毛虫lgG抗体的ELISA方法.当采集血清标本有困难时,可将唾液替代血清用于检测旋毛虫病.     

日本血吸虫不同抗原检测家兔感染及治疗前后IgG/IgM抗体动态

目的探索特异性抗体检测在日本血吸虫病诊断和疗效考核中的潜在应用价值。方法ELISA方法采用可溶性虫卵抗原(SEA)、成虫抗原(AWA)检测人工感染家兔治疗前后血清中特异性IgGI、gM抗体,评价特异性抗体检测的诊断和疗效考核价值。结果用SEA抗原检测IgM抗体,发现感染后7周不管治疗与否,抗体水平均快速下降。与SEA相比,感染后AWA特异性IgM抗体上升时间早,且治疗后5个月仍为阳性。SEA特异性IgG抗体水平最高,但治疗后5个月仍为阳性。结论采用SEA抗原检测IgG用于血吸虫病诊断效果较好;对早期感染的诊断可以考虑SEA抗原检测IgM抗体作辅助。AWA抗原检测IgM,对急、慢性血吸虫感染均有较好诊断效果。采用这两种抗原检测IgG和IgM,均无考核疗效意义。     

Varicella-Zoster Immunoglobulins during Varicella, Latency, and Zoster.

Antibodies to membrane antigens of cells infected by varicella-zoster virus were detected by immunofluorescence. Rises in varicella-zoster IgA, IgG, and tigM were detected after both varicella and zoster. Prompt antibody responses were observed in patients with generalized zoster as well as in those with localized zoster. A delayed response was found, moreover, in one patient who did not develop disseminated disease. Antibody to varicella-zoster virus was detected in the sera of all three patients tested prior to and in all but one of 63 patients tested soon after onset of zoster. The level of varicella-zoster IgA and IgG rose in two of three immune patients after household exposure to infection with varicella-zoster virus. Varicella-zoster IgG was detected in the cord blood of infants whose mothers were immune to varicella.