Development and Clinical Application of a Single-tube Nested PCR Method to Amplify the DNA Polymerase Ⅰ Gene of Treponema Pallidum

Objective: To develop a sensitive, specific and simple method for detection of extremely low numbers of T. pallidum in clinical specimens, as a significant addition to the serologic tests for syphilis diagnosis. Methods: Double-tube nested PCR(DN-PCR) and single-tube nested PCR(SN-PCR) assays were performed to amplify specific fragments of the DNA poly-merase I gene(polA) of T. pallidum. Sensitivity and specificity of the two PCR assays were tested. Eighty-six whole blood specimens from persons with suspected syphilis were detected by the two nested PCR methods. The TPPA test was used as a comparison for detecting syphilis in sera from corresponding patients. Results: Only specific amplicons could be obtained during amplification of the T. pallidum polA gene and the detection limit was approximately 1 organism when analyzed on gel by the two PCR methods. Of 86 clinical specimens, 62 were positive by TPPA. Of these, 54 and 51 were positive by the DN-PCR and SN-PCR, respectively, which does not represent a statistically significant difference between the two PCR tests. Of 24 TPPA-negative specimens, 5 were positive by both DN-PCR assay and SN-PCR assay. Conclusion: The SN- polA PCR method is extremely sensitive, specific and easy to perform for detecting low numbers of T. pallidum in clinical blood specimens as a complementary to serology for syphilis diagnosis.

Construction and Expression of Tp0453 Recombinant Protein of Treponema pallidum and Development of Indirect ELISA for Diagnosinq Syphilis

Objectives： To clone and express Tp0453 outer membrane protein of Treponema pallidum, and to evaluate its significance in the serodiagnosis of syphilis. Methods： The immuno-dominant epitope of Tp0453 gene was amplified from the complete genome of T.pallidum by polymerase chain reactions （PCR）, subcloned into the expression vector Pqe32 to generate recombinant plasmid Pqe32/Tp0453, and was then expressed in E. coli M15. The fusion protein was purified with Ni-NTA affinity purification. Indirect ELISA was developed to detect human serum IgG antibody to T. pallidum. Results： The recombinant Tp0453 protein was successfully expressed and purified. The recombinant protein had a molecular weight of approximately 32KDa.Indirect ELISA to the recombinant protein was developed.Sixty control sera were tested by ELISA and yielded a sensitivity of 100% （30/30） and a specificity of 100% （30/30）. While testing for T. pallidum in human sera, the sensitivity and specificity of ELISA were 96.8% and 100%, respectively, when compared with TPPA test results. The concordance of results between the ELISA test and the TPPA test was 98.2%. Conclusion： The recombinant Tp0453 outer membrane protein elicited a strong immunoreaction to anti-T.pallidum IgG antibody and has great potential use in ELISA for the serodiagnosis of syphilis.

Treponema pallidum and Haemophilus ducreyi DNA detection by A Multi-Nested Polymerase Chain Reaction

Objectives: To develop a multi-nested polymerase chain reaction in an assay to detect early Treponema pallidum and Haemophilius ducreyi DNA in the swabs of genital ulcers. Methods: Four pairs of outer and inner primers, specific to the basic membrane protein gene of Treponema pallidum and to the 16s rRNA gene of H ducreyi were synthesized. The multi-nested PCR was developed and applied to detect Treponema pallidum and Haemophilus dicreyi in clinical swabs. Result: The two samples of standard strains of Haemophilus ducreyi and one Treponema pallidum were amplified and showed 309-bp rRNA gene of Haemophilus ducreyi and 506-bp DNA of Treponema palidum, respectively. Out of 51 samples of genital ulcer detected, 29 showed Treponema pallidum positive product and no Haemophilus ducreyi DNA was found. Conclusion: The multi-nested PCR for Treponema pallidum and Haemophilus ducreyi could be useful for early detection and distinguishing diagnosis between syphilis and chancroid.

CLINICAL EVALUATION OF FOUR RECOMBINANT TREPONEMA PALLIDUM ANTIGEN-BASED RAPID TESTS IN THE DIAGNOSIS OF SYPHILIS

Objective To assess the sensitivity, specificity, and feasibility of 4 recombinant Treponema pallidum antigenbased rapid tests in the diagnosis of syphilis. Methods A total of 970 outpatients were selected from the Sexually Transmitted Diseases Centre of Peking Union Medical College Hospital. Venous blood was collected and serum was extracted. T. paUidum antibodies in whole blood, anticoagulant whole blood, and serum were detected using 4 recombinant T. pallidum antigen-based rapid tests. T. pallidum haemagglutination test （TPHA） was considered as the gold standard for the detection of T. pallidum specific antibodies in serum. The sensitivities and specificities of four methods were analyzed. Results The sensitivities and specificities of Abbott Determine Syphilis TP test, SD-BIOLINE Syphilis 3.0 test, VISITECT-SYPHILIS test, and Syphicheck-WB test for serum specimens were 100% and 98. 9%, 95.7% and 98.0%, 94.6% and 98.2%, 68.1% and 98.9% ; for whole blood were 74. 1% and 99. 5%, 87.9% and 99.4% , 73.2% and 99.7%, 64. 7% and 99.7%. The observed sensitivities of the 4 rapid diagnosis tests were not significantly different with TPHA （ P 〉 0.05 ）. Conclusions The 4 rapid tests show good performance and characteristics in the diagnosis of syphilis. Furthermore, they are more sensitive for serum specimens than whole blood.

Construction of the Eukaryotic Expression Vector for Outer Membrane Protein Tp92 from Treponema pallidum and Its Preliminary Study on the Immune Responses in New Zealand Rabbits

To construct the recombinant plasmid of eukaryotic expression containing Tp92 gene from Treponema pallidum and study its immunogenicity in New Zealand white rabbits. Tp92 gene was amplified from the genomic DNA of T. pallidum by polymerase chain reaction (PCR) and subcloned into appropriate site of pcDNA3.1(+) vector. After identification by sequencing and restrictive enzyme digestion, the recombinant plasmid was transfected into HeLa cells using liposome, and the expressed protein was identified by immunocytochemistry and Western blotting. After verifying that the Tp92 antigen gene fragment could be expressed in HeLa cells, 100?μg of recombinant plasmids [pcDNA3.1(+)-Tp92], 100 μg of control plasmids [pcDNA3.1(+)] or 0.5 ml PBS buffer were administered in 3 groups of New Zealand white rabbits (6 rabbits/group), and the booster immunizations were employed at 2-week interval for 3 times. ELISA assay was used for the quantitative detection of the specific antibody in the sera of rabbits, and the proliferation response of spleen cells was detected by MTT assay. It was found that the target gene Tp92 segment about 2103 bp was obtained, and the DNA sequence of Tp92 gene constructed in pcDNA3.1 (+) vector was consistent with the published nucleotide sequence. The homologies of the nucleotide and putative amino acid sequences of Tp92 gene between T.pallidum subsp. pallidum Nichols and various pathogenic treponeme strains were 95.5%-100%. The analysis of immunocytochemistry and Western blotting showed that Tp92 gene segment constructed in pcDNA3.1(+) vector could express a fusion protein with a calculated molecular mass of 77 kDa in HeLa cells and the expressed protein could react with positive blood serum from syphilis patient. The specific antibody IgG titers were observed and the highest titer was 1∶1024 in rabbits after 3 times with pcDNA3.1(+)-Tp92. The proliferation response of spleen cells were significantly higher than that of rabbits injected with pcDNA3.1(+) ( P <0.05). The successful expression of the eukaryotic expression plasmid of Tp92 gene from T. pallidum was obtained in eukaryotic system and strong responses of humoral and cellular immunity was evoked by DNA vaccine of pcDNA3.1(+)-Tp92 in rabbits thus establishing a solid basis for the future studies in the biological activities and for the development of the syphilis DNA vaccine.

Expression and purification of the recombinant outermembrane protein Tp0453 of Treponema pallidum and its characterization of immuno-competence

To clone and express the recombinant outer membrane protein Tp0453 of Treponema pallidum and to analyze the immuno-reactivity and immunogenicity of the expressed protein, the immuno-dominant epitope of the Tp0453 was amplified from the complete genome of T.pallidum by PCR, subcloned into expression vector pQE32 to generate the recombinant plasmid pQE32/Tp0453, then expressed in E.coli M15 and analyzed by SDS/PAGE and Western blotting. The fusion protein expressed was purified with Ni-NTA affinity chromatography. Its immuno-reactivity was assayed by indirect ELISA, and the immunogenicity was determined by immunization with this fusion protein in New Zealand rabbits. In the present study, a fusion protein of molecular weight about 32 kDa was obtained. As demonstrated by Western blotting, the recombinant protein could react specifically with positive IgG sera of patients with syphilis, and the antibodies against T.pallidum in human sera were successfully detected by indirect ELISA. Both the sensitivity and specificity of ELISA based on the Tp0453 fusion protein as were 100% (30/30) when detected with control sera. In comparison with the results of IgG ELISA with those of TPPA. It was found that the sensitivity of ELISA was 96.8% and the specificity was 100%. The difference of ELISA and TPPA was not significant, and the concordance of results between ELISA and TPPA was 98.2%. In addition, specific humoral responses could be elicited by immunization with the recombinant fusion protein in New Zealand rabbits with a specific antibody titer of 1∶1280 after 3 successive doses of immunization. These results demonstrate that the expressed recombinant fusion protein shows excellent immuno-competence and provide foundation to develop a quick diagnostic kid applied to detect the presence of T.pallidum infections.

Eukaryotic expression of outer membrane protein Gpd from Treponema pallidum and preliminary studies on its immune response in rabbits

The Gpd gene was amplified from the genomic DNA of Treponema the appropriate site of pcDNA3.1 （ ＋ ） vector. The expression of pcDNA3. I was tested with Western blotting and technology of immunoeytochemisty. New munized with the eukaryotic expression recombinant pcDNA3, 1 （ ＋ ）-Gpd pallidum and cloned into （ ＋ ）-Gpd in Hel,a cells Zealand rabbits were imA fusion protein of C, pd with 4.1 kDa has been effectively expressed in HeLa cells, which were detected bv Western blotting and the immunocytochemistry techniques. The New Zealand rabbits were able to elicit the specific antibody after immunization with the nucleic acid vaccine. The antibody titer could reach as high as 1 ： 1024 after 2 weeks of the third injection; and the splenocytes proliferated evidently due to the Gpd protein stimulation. Both the antibody titer and the splenocytes proliferation were higher substantially than those of controls （ P 〈 0.01 ）. All above data will contribute to an experimental basis of further study of the biological function of Gpd protein as well as DNA vaccine for syphilis.

Detection of Treponema pallidum,Herpes Simplex Virus,and Haemophilus ducreyi from Genital Ulcers by Multiples Polymerase Chain Reaction

Objective: To evaluate the clinical application of multiplex PCR inthe detection of Treponema pallidum, Herpes simplex virus (HSV), andHaemophilus ducreyi. Method: Three standard strains were used to set up a multiplexPCR (MPCR) for detecting syphilis, herpes genitalis, and chancroidsimultaneously. Samples from 122 patients with genital ulcer disease(GUD) were subjected to MPCR and the results were compared withthose of dark-field microscopy and TP serology, HSV antigen ELISA,and H. ducreyi culture. Results: In the 122 patients with GUD, MPCR identified 34 casesof T. pallidum infection, 40 cases of HSV infection, and 2 cases of mixedinfection of T. pollidum and herpes. No positive results of H. ducreyiwere found. The sensitivity of MPCR to T. pallidum and herpes was100% and 93.3%, respectively. The sensitivities of dark-field mi-croscopy and TP serology, HSV antigen ELISA, and H. ducreyi cul-ture was 35.3%, 50% and 100%, respectively. Conclusion: MPCR showed a relatively higher sensitivity for T.pallidum as compared with the routine techniques. Although its sensi-tivity for HSV was not as good as that of antigen ELISA, it also yieldeda high detection rate. MPCR can detect more than one pathogen. It issimple, quick, sensitive, and suitable for clinical use or epidemiologicalinvestigation.

The Construction of the Eukaryotic Expression Vector of Glycerophosphodiester Phosphodiesterase Gene from Treponema pallidum and its Expression in Hela Cells

Objective: To construct the recombinant plasmid containing Glycerophosphodiester phosphodiesterase (Gpd) gene from Treponema pallidum and transfect it into Hela cells to express the encoded outer membrane protein. Methods: The Gpd gene was amplified from the genomic DNA of T.pallidum by polymerase chain reaction (PCR) and inserted into cloning vector pUCm-T. The inserted Gpd gene was subcloned into the appropriate site of pcDNA3.1(+) vector. After identification by sequencing and restrictive enzymes digestion, the recombinant plasmid was transfected into Hela cells using liposomes. The expressed protein was identified by immunocytochemistry and Western blot. Results: The target Gpd gene segment was approximately 1059bp. The DNA sequence of the Gpd gene contained in the pcDNA3.1(+) vector was consistent with the published nucleotide sequence. The homology of the nucleotide and putative amino acid sequences of the Gpd gene between T. pallidum subsp. pallidum Nichols and various pathogenic treponemal strains ranged from 98% to 100%. Immunocytochemistry and Western blot analysis showed that the constructed Gpd-pcDNA3.1(+) vector expressed a fusion protein with a calculated molecular mass of 41KDa in Hela cells and that the expressed protein reacted with the sera from syphilis patients. Conclusion: The successful construction and expression of the eukaryotic expression plasmid of the Gpd gene from T.pallidum provide a promising tool to further study the biological activity of T.pallidum and develop a DNA vaccine for syphilis.

Cloning and Expression of the Tpp17 Gene of Treponema pallidum And Clinical Application

Objective: To obtain recombinant Treponema pallidumsubsp pallidum (TP 17KD) lipoprotein in large quantities byamplification and to further purify antigens for laboratorydiagnosis of syphilis and development of a syphilis vaccine. Method: The Tpp17 lipoprotein gene was amplified fromthe TP(strain Nichols), and then it was recombinated into aplasmid pMAL-2c and cloned within E coli l2-TB1. The hostbacteria containing recombinant plasmids were induced withIPTG. The Tpp 17KD lipoprotein gene was amplified by us-ing PCR and positive clones were screened with double diges-tion and PCR. Recombinant plasmids were transformed intoE. coli and the E coli carrying recombinant plasmids wereinduced. The expression of TP 17KD was detected by sodiumdedecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and immunoblot. Results:Gel staining with Coomassie blue G-250 showedthat the induced E coli carrying recombinant plasmid couldproduce 60KD fusion protein at high levels. Gel scanningshowed that 17KD protein expression in E coli accounted for10% of total cellular protein. The recombinant protein antigenreacted with the sera of syphilis patients. Conclusion: Our study lays a cornerstone for developingnew techniques of laboratory diagnosis for syphilis and newvaccines. Preliminary clinical application showed that thefusion protein could be used for the diagnosis of syphilis.

Treponema pallidum-specific antibody expression for the diagnosis of different stages of syphilis

Background Tp15, Tp17, Tp45, and Tp47 are outer-membrane proteins found in Treponema pallidum, the etiologic agent of syphilis. These proteins are potent antigens and are potential markers for the serological detection of syphilis. The present study analyzed antibodies to these protein antigens （TP-IgM and TP-IgG） in human serum and investigated the expression of these antibodies during different stages of syphilis. Methods Serum samples were collected from 69 subjects （male 45, female 24） diagnosed with syphilis and analyzed by Western blotting for the expression of IgM and IgG against the four protein antigens. Expression levels of the target antibodies were compared during the same stage of syphilis as well as between different stages of this disease. Results In subjects with primary syphilis, the positive rate of Tp45 IgM was higher than that of other TP-IgM. Tp15 IgM was detected only in subjects with tertiary syphilis. Similarly, the seroprevalence of Tp45 IgG in primary syphilis was higher than for other TP-IgG. No target TP-IgM was detected in subjects with latent syphilis. In subjects with secondary syphilis, the expression level of Tp15 IgG （138.73±20.16） was higher than for other target TP-IgG. In subjects with tertiary syphilis, all target TP-IgG were detected. In subjects with tertiary or latent syphilis, the expression levels of Tp45 IgG （121.33±11.04 and 110.10±40.19, respectively） were higher than those of other target TP-IgG. The expression levels of all Tp-lgM were similar before or after anti-syphilis treatment. In comparison, the expression levels of all TP-IgG decreased compared with the pre-treatment levels, and this decrease was statistically significant （both P 〈0.05） for Tp17 IgG and Tp47 IgG. Conclusions After Treponema pallidum infection, Tp45 IgM appeared first and Tp15 IgM occurred during later stages. The positive rates of all TP-IgG increased with the duration of this disease. Anti-syphilis treatment reduced the expression levels of Tp17 IgG and Tp47 IgG. Larger-scale studies are required to further validate the value of Tp15, Tp17, Tp45, and Tp47 as markers for the early detection of primary and latent syphilis.

Seroreactivity and immunogenicity of Tp0965, a hypothetical membrane protein of Treponema pallidum

Background Treponema pallidum （T. pallidum） subsp, pallidum is the causative agent of syphilis. Analysis of recombinant antigens of T. pallidum led to the identification of potential candidate antigens for vaccine development and syphilis serodiagnosis. Tp0965 was predicted to be a membrane fusion protein and was found to be reactive with infected human sera in previous studies, but the results were controversial. In this research, the antigenicity and immunoreactivity of recombinant protein Tp0965 were assessed. Methods T. pallidum subsp, pallidum （Nichols strain） was propagated and isolated and the genomic DNA was extracted. The Tp0965 gene was amplified by polymerase chain reaction （PCR）. Then the recombinant protein Tp0965 was expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid （Ni-NTA） purification system. The reactivities of protein Tp0965 were examined by immunoblot analysis and indirect enzyme-linked immunosorbent assay. The antisera against protein Tp0965 were obtained by immune rabbits and the immunogenicity of antisera were detected by indirect enzyme-linked immunosorbent assay. Results Recombinant protein Tp0965 was expressed successfully in vitro. Immunoblot assay showed that the recombinant protein Tp0965 could be recognized by human syphilitic sera of all stages. Indirect enzyme-linked immunosorbent assay showed there were only 4 of 74 human syphilitic sera that failed to show reactivity to recombinant antigen Tp0965, and lack of reactivity of Tp0965 to all 28 uninfected sera. A low titer of antiserum against Tp0965 in immune rabbits could be detected after the third time of immunization. Conclusions The recombinant antigen Tp0965 shows excellent sensitivity for the reactivity with sera from syphilitic individuals at all stages. The results also demonstrate a potential application for the serodiagnosis of syphilis.

Production of proinflammatory cytokines in the human THP-1 monocyte cell line following induction by Tp0751,a recombinant protein of Treponema pallidum

The tissue destruction characteristic of syphilis infection may be caused by inflammation due to Treponema pallidum and the ensuing immune responses to the pathogen.T.pallidum membrane proteins are thought to be potent inducers of inflammation during the early stages of infection.However,the actual membrane proteins that induce inflammatory cytokine production are not known,nor are the molecular mechanisms responsible for triggering and sustaining the inflammatory cascades.In the present study,Tp0751 recombinant protein from T.pallidum was found to induce the production of proinflammatory cytokines,including TNF-α,IL-1βand IL-6,in a THP-1 human monocyte cell line.The signal transduction pathways involved in the production of these cytokines were then further investigated.No inhibition of TNF-a,IL-1β,or IL-6 production was observed following treatment with the SAPK/JNK specific inhibitor SP600125 or with an ERK inhibitor PD98059.By contrast,anti-TLR2 mAb,anti-CD14 mAb,and the p38 inhibitor SB203580 significantly inhibited the production of all three cytokines.In addition,pyrrolidine dithiocarbamate (PDTC),a specific inhibitor of NF-κB,profoundly inhibited the production of these cytokines.Tp0751 treatment strongly activated NF-κB,as revealed by Western blotting.However,NF-κB translocation was significantly inhibited by treatment with PDTC.These results indicated that TLR2,CD14,MAPKs/p38,and NF-κB might be implicated in the inflammatory reaction caused by T.pallidum infection.

梅毒实验室诊断的研究现状与展望

[背景]近年来,梅毒发病数呈逐年上升趋势,引起广泛关注,已成为全球性公共卫生问题.鉴于梅毒病程的隐匿性、复杂性和多样性,迫切需要高灵敏度的诊断方法和有效的疗效监测指标,以实现梅毒的早诊断和早治疗.[进展]本文对梅毒实验室检测方法及诊断程序的研究现状进行综述.病原学检查可以直接确诊感染,但易受取材部位及取材方法等因素影响,灵敏度较低.血清学检测是梅毒诊断的主要检测方法,但由于抗体至少在感染2周后才产生,难以筛查窗口期的患者.由于血清学诊断程序的行业标准尚未统一,各实验室对3种血清学诊断程序的选择也不尽相同.聚合酶链式反应(polymerase chain reaction,PCR)对于一期或二期梅毒患者皮损组织具有较高灵敏度和特异度,但对于较易获得的血液样本,其梅毒螺旋体(Treponema pallidum)DNA检出率低,国内至今尚无获得医疗器械注册证的梅毒PCR检测试剂,无法应用于临床检测.[展望]首先,研发梅毒螺旋体抗体和非特异性抗体联合检测试剂,避免单一试验假阴性造成的漏诊;其次,开展血液、脑脊液、尿液等标本的梅毒螺旋体抗原谱研究,缩短检测的窗口期;最后,引入质量管理体系,制定梅毒分子诊断的统一标准.

活动性梅毒患者梅毒血清特异性抗体水平与体内免疫球蛋白水平的相关性研究

目的:探究活动性梅毒患者梅毒血清特异性抗体水平(TPAb)与体内免疫球蛋白A/E/M/G(IgA/E/M/G)及IgG亚型的相关性,为临床梅毒的综合诊疗与预后评估提供依据。方法:选取2023年就诊于苏州大学附属第一医院的活动性梅毒患者,根据梅毒甲苯胺红不加热血清试验(TRUST)分为1∶4+与1∶16+两组,另选取健康对照组,均进行化学发光微粒子免疫检测(CMIA)测定TPAb,免疫比浊法测定IgA/E/M/G及IgG1/2/3/4含量。结果:本次共纳入活动性梅毒患者206例(男性125例,女性81例),年龄(46.05±19.58)岁。相对健康对照组,可见活动性梅毒组IgG2降低,TPAb、IgA/E/M/G和IgG1/3/4水平均升高,其中TPAb、IgE、IgG1/2/3水平差异具有统计学意义(P<0.05)。相比于健康对照组,TRUST 1∶4+组TPAb、IgE、IgG2差异具有统计学意义,TPAb与IgE、IgG2/4存在相关性;而TRUST 1∶16+组TPAb、IgA/E/M/G、IgG1/2/3水平差异均具有统计学意义,TPAb与IgE、IgG4存在相关性(P<0.05)。另外,TRUST阳性组之间的TPAb、IgG和IgG1/3水平差异具有统计学意义(P<0.05)。结论:不同TRUST滴度的活动性梅毒患者TPAb与Ig水平存在较大差异,TPAb联合Ig可用于监测梅毒活动性强弱,IgA/M/G及IgG1/3可能成为候选生物标志物。

抗-TP阳性献血者中抗病毒治疗药物使用情况分析

目的了解深圳市梅毒螺旋体(TP)抗体(抗-TP)阳性献血者中抗逆转录病毒治疗(ART)药物使用情况,评估人类免疫缺陷病毒(HIV)诊疗新趋势带来的血液安全风险。方法采用分层随机抽样方法选取2019年3月至2023年1月深圳地区血液筛查合格的重复献血者60例(阴性对照组),规律服用已知ART药物人群3例(阳性对照组),抗-TP阳性/抗-HIV阴性献血者353例(实验1组),抗-TP阳性/抗-HIV阳性献血者25例(实验2组),应用高效液相色谱-串联质谱(HPLC-MS/MS)方法检测各组血浆样本中8种ART药物浓度,分析ART药物使用情况。结果阳性对照组血浆采用1∶6稀释混样后ART药物仍可检出,实验1组和实验2组1∶6人份混合血浆阳性样本经拆分确证,实验2组检出1例ART药物阳性样本,该样本抗-HIV、蛋白免疫印迹、HIV RNA阳性,抗TP阳性献血者ART药物检出率为0.26%,实验1组ART药物检出率为0.00%,实验2组ART药物检出率为4.00%。结论深圳地区抗-TP阳性献血者中发现使用ART药物情况,合并HIV感染及高危性行为人群更可能使用ART药物。

2019—2023年江西省血液中心无偿献血者人类免疫缺陷病毒及梅毒螺旋体感染现状及特征分析

目的:调查2019—2023年江西省血液中心无偿献血者人类免疫缺陷病毒(HIV)及梅毒螺旋体(TP)感染现状,并分析其特征。方法:选取2019年1月—2023年12月江西省血液中心的408605名无偿献血者血液标本进行调查,对标本进行HIV、TP项目常规检测,HIV反应性标本送HIV确证实验室进行确证。结果:2019—2023年无偿献血者HIV初筛阳性率为0.062%,TP初筛阳性率为0.227%,总阳性率为0.289%。各年度初筛总阳性率比较,差异有统计学意义(P<0.05)。免疫印迹法确证检测结果显示,34名无偿献血者为HIV阳性,HIV阳性率为0.008%。2019—2023年的无偿献血者中,女性HIV及TP初筛阳性率(0.324%)高于男性(0.268%),45~<56岁的无偿献血者HIV及TP初筛阳性率(0.542%)较高,研究生HIV及TP初筛阳性率(0.160%)较低,初筛阳性率随学历降低而呈现升高的趋势,农民HIV及TP初筛阳性率(0.629%)较高,初次献血者HIV及TP初筛阳性率(0.417%)较高。2019—2023年34名HIV确证阳性的献血者中,女性占比(5.882%)低于男性(94.118%),18~<25岁者占比(50.000%)最高,学生占比(41.176%)最高。结论:不同特征的无偿献血者HIV及TP检测结果存在差异,可通过加强健康教育和“献血后回告”宣教、强化献血前征询与筛查、构建区域性的献血者监测系统和艾滋病疫情管理共享系统等方式推进我国血液安全。

Infectious Disease Update in Obstetrics: A Modern Approach to the Patient with a Positive Screening Test for Syphyilis in Pregnancy

Screening for maternal syphilis has been an essential component of routine antenatal screening tests in most countries for many years. This is not only because of the virulence of the spirochete which causes the infection but also because of its vertical transmission rate and the potential severe adverse complications/morbidity that can result from its transmission to the fetus. Although the incidence of maternal syphilis and its fetal sequalae in low-income countries has been considerable for several years, the disease has been almost non-existent in high income countries with wide antenatal screening coverage and effective treatment programmes for Syphilis. The recent alarming increase in the incidence of maternal syphilis in high income countries has spawned a renewed public health interest in the infection, with several countries updating and strengthening public health guidance in an attempt to stem this dramatic trend. This is a short clinical update for the practising obstetrician on how to manage the antenatal patient with a positive syphilis screening test.

2018-2022年北京市密云区医院孕产妇人类免疫缺陷病毒、梅毒螺旋体、乙型肝炎病毒检测结果分析

目的:分析2018-2022年北京市密云区医院孕产妇人类免疫缺陷病毒(HIV)、梅毒螺旋体、乙型肝炎病毒(HBV)检测结果。方法:选取2018年1月-2022年12月于密云区医院进行孕产期保健及分娩的孕产妇19 327例作为研究对象,收集并分析孕产妇一般资料及HIV、梅毒螺旋体和HBV初筛结果。结果:19 327例孕产妇中,HIV阳性0例,梅毒螺旋体阳性101例(0.52%),HBV阳性175例(0.91%)。2018-2022年梅毒螺旋体和HBV阳性呈小幅度波动,其中以2021年明显降低。孕产妇梅毒螺旋体、HBV阳性率及构成比均以20~29岁最高,40~45岁最低。结论:2018-2022年北京市密云区医院孕产妇HIV、梅毒螺旋体、HBV阳性率较低,以20~29岁最多,临床应针对此情况采取应对措施,消除艾滋病、梅毒、乙型肝炎的母婴传播。

探讨TRUST与TP-ELISA对梅毒患者的诊断价值

目的 探讨甲苯胺红不加热血清试验(toludine red unheated serum test, TRUST)、梅毒螺旋体-酶联免疫吸附试验(treponema pallidum-enzyme linked immunosorbent assay, TP-ELISA)检测方法在梅毒诊断中的诊断价值。方法 非随机选取2022年1月—2023年10月兴义市人民医院收治的86例疑似梅毒患者为研究对象,所有患者均完成TRUST与TP-ELISA检测,并将梅毒螺旋体明胶颗粒凝集试验(treponema palli-dum particle agglutination, TPPA)检测作为金标准,分析TRUST与TP-ELISA的诊断价值。结果 86例疑诊梅毒患者中,61例患者确诊为梅毒,患病率为70.93%(61/86)。TRUST检查的阳性率为59.30%(51/86),阴性率为40.70%(35/86);TP-ELISA检查的阳性率为67.44%(58/86),阳性率为32.56%(28/86)。TP-ELISA的敏感度为93.44%(57/61),高于TRUST检查的80.33%(49/61),差异有统计学意义(χ^(2)=4.604,P=0.032);且TP-ELISA准确度为94.19%(81/86),高于TRUST的83.72%(72/86),差异有统计学意义(χ^(2)=4.793,P=0.029)。结论 在梅毒诊断中,TP-ELISA的敏感度、准确度高于TRUST,但筛查过程仍存在假阴性风险,临床应综合考虑患者的危险因素筛查结果与其他临床指标。