Objective: The aim of the present study was to evaluate the performance of the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA using one dried blood spot (DBS) as an alternative sample to plasma. Method: A total of 571 paired DBS/plasma samples were collected from men who have sex with men (MSM) and injection drug users (IDUs), and serological and molecular assays were performed. Using plasma results as the reference standard, the performance of DBS tests for HIV-1 RNA, HIV-1 DNA, and HCV RNA was evaluated. Pearson's correlation coefficients and Bland-Altman analysis were performed to assess the correlation and concordance between DBS and plasma. Results: Among paired plasma/DBS samples with detectable HIV-1 RNA and HCV RNA, five samples (5/32) were not detectable in DBS, while measurable HIV-1 RNA levels were present in plasma (1.44 to 3.99 log Conclusion The performance of the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA using one DBS was acceptable. DBS, as an alternative sample to plasma, may be a viable option for the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA in resource-limited settings or for individuals living in areas that are difficult to access.
DNA, Viral/analysis* ; Diagnostic Tests, Routine/methods* ; Dried Blood Spot Testing/methods* ; HIV Infections/diagnosis* ; HIV-1/isolation & purification* ; Hepacivirus/isolation & purification* ; Hepatitis C/diagnosis* ; RNA, Viral/analysis* ; Sensitivity and Specificity ; Specimen Handling/methods* ; Syphilis/diagnosis* ; Treponema pallidum/isolation & purification*

BACKGROUND: Automated Mediace Treponema pallidum latex agglutination (TPLA) and Mediace rapid plasma reagin (RPR) assays are used by many laboratories for syphilis diagnosis. This study compared the results of the traditional syphilis screening algorithm and a reverse algorithm using automated Mediace RPR or Mediace TPLA as first-line screening assays in subjects undergoing a health checkup. METHODS: Samples from 24,681 persons were included in this study. We routinely performed Mediace RPR and Mediace TPLA simultaneously. Results were analyzed according to both the traditional algorithm and reverse algorithm. Samples with discordant results on the reverse algorithm (e.g., positive Mediace TPLA, negative Mediace RPR) were tested with Treponema pallidum particle agglutination (TPPA). RESULTS: Among the 24,681 samples, 30 (0.1%) were found positive by traditional screening, and 190 (0.8%) by reverse screening. The identified syphilis rate and overall false-positive rate according to the traditional algorithm were lower than those according to the reverse algorithm (0.07% and 0.05% vs. 0.64% and 0.13%, respectively). A total of 173 discordant samples were tested with TPPA by using the reverse algorithm, of which 140 (80.9%) were TPPA positive. CONCLUSIONS: Despite the increased false-positive results in populations with a low prevalence of syphilis, the reverse algorithm detected 140 samples with treponemal antibody that went undetected by the traditional algorithm. The reverse algorithm using Mediace TPLA as a screening test is more sensitive for the detection of syphilis.
Algorithms ; Anti-Bacterial Agents/therapeutic use ; Humans ; Latex Fixation Tests ; Reagins/blood ; Syphilis/*diagnosis/drug therapy/microbiology ; Treponema pallidum/isolation & purification

OBJECTIVETo analyze the prevalence rates and associated risk factors on HIV, syphilis, herpes simplex virus-2 (HSV-2) among female sex workers (FSWs) in Jiaozhou, Shandong province.

METHODSThrough convenient sampling, an anonymous questionnaire survey was conducted on female sex workers to collect related information. Blood specimens were drawn for serological tests on HIV, syphilis and HSV-2 antibodies, respectively. Patients with positive results from both treponema pallidum and HSV-2 tested by ELISA method, were defined as being superinfected.

RESULTSA total of 460 FSWs were recruited in this study. The prevalence rates of HIV, syphilis, HSV-2, and syphilis/HSV-2 superinfection were 0.22% , 5.9% , 43.0% , and 11.7% , respectively. Among the methamphetamine users, the prevalence rates of syphilis, HSV-2, and syphilis/HSV-2 superinfection were 12.4%, 55.2%, and 22.9%, respectively. Results from multivariate analysis showed that factors that significantly associated with syphilis infection would include: methamphetamine use (OR = 3.61, 95% CI:1.62-8.06), having first commercial sex intercourse at under 20 years of age (OR = 2.80, 95% CI:1.15-6.85), more than 2 establishments that the FSWs worked in the last month (OR = 4.37, 95%CI:0.83-22.83). Factors associated with HSV-2 infection were methamphetamine use (OR = 2.30, 95%CI:1.43-3.70), having first commercial sex intercourse at under 20 years of age (OR = 1.77, 95% CI:1.16-2.69), working at low-end establishments (OR = 2.61, 95%CI:1.66-4.10), working at the local sex-work venues for more than one year (OR = 2.01, 95% CI:1.28-3.14), with low education backgrounds (OR = 2.27, 95% CI:1.18-4.36), using methamphetamine (OR = 3.95, 95% CI:2.09-7.44), low education background (OR = 2.43, 95% CI: 1.10-5.36), 21-30 year-olds (OR = 2.95, 95%CI:1.08-8.03), older than 31 years of age (OR = 7.05, 95% CI:2.48-20.01) etc., were independent risk factors associated with the superinfection of syphilis and HSV-2.

CONCLUSIONThe prevalence rates of syphilis, HSV-2 and syphilis/HSV-2 superinfection were relatively high among FSWs in Jiaozhou, especially among the FSWs who were methamphetamine users.

Age Distribution ; Amphetamine-Related Disorders ; epidemiology ; China ; epidemiology ; Educational Status ; Female ; HIV Infections ; epidemiology ; HIV Seropositivity ; Herpes Genitalis ; epidemiology ; Herpesvirus 2, Human ; isolation & purification ; Humans ; Methamphetamine ; administration & dosage ; Prevalence ; Risk Factors ; Serologic Tests ; Sex Work ; Sex Workers ; psychology ; statistics & numerical data ; Superinfection ; epidemiology ; Syphilis ; epidemiology ; Treponema pallidum ; isolation & purification

Syphilis, a chronic bacterial infection caused by the spirochete Treponema pallidum subsp. pallidum, remains a public health concern worldwide. Syphilis control is largely dependent upon early identification and prompt treatment. The diagnosis of syphilis is mainly based on laboratory tests, especially serology and dark-field microscopy. In recent years, recombinant Treponema pallidum antigen-based rapid tests, polymerase chain reaction, and immunoglobulin M antibody detection have also shown certain sensitivities and specificities for syphilitic patients at different stages.
Antigens, Bacterial ; blood ; Humans ; Immunoglobulin M ; blood ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Syphilis ; blood ; diagnosis ; Syphilis Serodiagnosis ; Treponema pallidum ; immunology ; isolation & purification

Adolescent ; Anti-Bacterial Agents ; therapeutic use ; Anti-Ulcer Agents ; therapeutic use ; Female ; Humans ; Penicillin G Procaine ; therapeutic use ; Stomach Ulcer ; drug therapy ; microbiology ; pathology ; Syphilis ; drug therapy ; microbiology ; pathology ; Syphilis Serodiagnosis ; methods ; Treponema pallidum ; isolation & purification

No abstract available.
Adult ; Duodenal Diseases/*diagnosis/microbiology/pathology ; Gastric Mucosa/microbiology/pathology ; Gastroscopy ; Humans ; Male ; Stomach Diseases/*diagnosis/microbiology/pathology ; Syphilis/*diagnosis/microbiology/pathology ; Tomography, X-Ray Computed ; Treponema pallidum/isolation & purification

OBJECTIVETo clone tpn17 and tpn47 genes of Treponema pallidum and then construct their prokaryotic expression systems,to establish ELISAs based on rTpN17 and rTpN47 as antigens and to evaluate the sensitivity and specificity of the ELISAs for detection of serological diagnosis of syphilis.

METHODSThe whole length of tpn17 and tpn47 genes was amplified by PCR and then their prokaryotic expression systems were constructed. SDS-PAGE was used to measure the expression of the target recombinant proteins rTpN17 and rTpN47. Ni-NTA affinity chromatography was applied to extract rTpN17 and rTpN47, while Western blot was performed to determine the specific immunoreactivity of rTpN17 and rTpN47. By using rTpN17 and rTpN47 as the coated antigen, respectively, ELISAs (rTpN17-ELISA and rTpN47-ELISA) were established to detect serum samples from 200 healthy individuals, 25 RA patients, 17 SLE patients and 211 syphilis patients. The detection effects of the ELISAs were compared to those of TRUST and TPHA.

RESULTThe sequence similarity of the cloned tpn17 and tpn47 genes was 100 % compared with the corresponding sequences in GenBank. The expression outputs of rTpN17 and rTpN47 were approximately 37.2 % and 26.8 % of the total bacterial proteins, respectively. Both the extracted rTpN17 and rTpN47 could take place remarkable conjugation reactions to the sera with positive antibody against Treponema pallidum.The positive detection rate of TPHA (99.1%) was the highest (P<0.001). The positive detection rates of rTpN17-ELISA (85.3 %) and rTpN47-ELISA (84.3 %) were similar (P>0.05). The positive detection rates of TRUST (72.5 %) was lower than that of rTpN17-ELISA (P=0.001) but similar to that of rTpN47-ELISA (P=0.014). The detection results of all the serum samples from healthy individuals, RA patients and SLE patients were negative, whereas 7.1 % (3/42) of the samples from RA or SLE patients were positive.

CONCLUSIONrTpN17 and rTpN47 are still maintaining their original immunoreactivity. The ELISAs using rTpN17 or rTpN47 as the antigen are rapid, simple and convenient, higher sensitivity and specificity methods for serological screening and detection of syphilis.

Antibodies, Bacterial ; Antigens, Bacterial ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Humans ; Male ; Syphilis ; diagnosis ; Syphilis Serodiagnosis ; Treponema pallidum ; chemistry ; immunology ; isolation & purification

BACKGROUND: We compared the results of automated and quantitative methods for the diagnosis of syphilis, Mediace Rapid Plasma Reagin (RPR) and Mediace Treponema pallidum Latex Agglutination (TPLA) (Sekisui Chemical Co., Ltd, Japan) with those of conventional methods. METHODS: Sera from 3,896 persons who had health checkups between December 2005 and November 2006 were included in the evaluation of positive rates and biological false positives (BFP) for Mediace RPR and TPLA. In addition, 134 patients' sera positive for automated Mediace RPR or TPLA were tested for VDRL and TPHA. Discrepancies between TPLA and TPHA results were confirmed by the RecomBlot Treponemal IgG/IgM (Mikrogen GmbH, Germany). Automated Mediace RPR and TPLA were performed using the Hitachi 7600 chemistry autoanalyzer (Hitachi, Japan). Samples with positive Mediace RPR and negative TPLA results were defined as BFP. RESULTS: Positive rate of automated Mediace RPR was 0.23% (9/3,896). BFP of the Mediace RPR was 0.18%. Positive rate of automated TPLA was 1.62% (37/2,284). Among the 134 patients' sera, 33 (24.6%) showed a discrepancy between conventional VDRL and automated Mediace RPR results: Among 31 Mediace RPR(+)/VDRL(-) sera, 13 were positive and 18 were negative for TPLA. The remaining 2 sera of discrepancy with Mediace RPR(-)/VDRL(+) were all positive for TPLA. There were seven sera that showed a discrepancy between automated TPLA and TPHA results: Two sera with Mediace RPR(+)/TPLA(-)/TPHA(+) showed negative recomBlot Treponemal IgG/IgM results, and among five sera with TPLA(+)/TPHA(-), three demonstrated IgG or IgM by recomBlot Treponemal IgG/IgM. CONCLUSIONS: The results of comparison data demonstrated that automated TPLA results had a high concordance with recomBlot Treponemal IgG/IgM results. Moreover, there are additional advantages of automated methods such as quantitative detection, low infection risk, and no influence by human handling.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Agglutination ; False Positive Reactions ; Female ; Humans ; Immunoglobulin G/analysis ; Immunoglobulin M/analysis ; Latex Fixation Tests ; Male ; Middle Aged ; Reagent Kits, Diagnostic ; Reagins/*blood ; Syphilis/*diagnosis ; Syphilis Serodiagnosis/*methods ; Treponema pallidum/*immunology/isolation & purification

OBJECTIVETo study the pathologic changes in nasal and oropharyngeal mucosa caused by treponema pallidum (TP) infection.

METHODSTwenty-five cases of nasal and oropharyngeal syphilis were retrieved from the archival files of Department of Pathology of Beijing Tong Ren Hospital collected during the period from June 1996 to September 2005. The hematoxylin and eosin-stained slides were reviewed. Histochemical study using modified Warthin-Starry stain and immunohistochemical study using polyclonal antibody for TP were carried out. The diagnosis of early syphilis was confirmed by rapid plasma regain (RPR) and TP hemagglutination (TPHA) tests.

RESULTSAmong the 25 cases studied, 20 showed neutrophil infiltration, microabscess formation and plasma cell infiltration in the lamina propria. Endothelial swelling of small blood vessels and syphilitic vasculitis was also seen. Tonsillar ulcers associated with abundant plasma cells, lymphocytes and histiocytes were noted in 14 cases. One of which demonstrated florid reactive lymphoid proliferation, with transforming lymphoid cells of various stages identified. Pseudoneoplastic squamous cell proliferation was seen in one case. Spirochetes were detected by modified Warthin-Starry stain in mucosal microabscesses and squamous epithelium in 20 cases, around small blood vessels in 5 cases, and on the surface of tonsillar ulcers in 14 cases. Abundant TP were also found in smears of exudates in 6 cases. TP antigen was detected in 4 cases by immunohistochemical staining. All the 25 cases studied were RPR (1:8 to 128) and TPHA-positive.

CONCLUSIONSEarly syphilis involving nasal cavity and oropharynx has distinctive pathologic features. Detailed histologic examination, together with modified Warthin-Starry stain for demonstration of spirochetes, is important to obtain a correct diagnosis.

Adult ; Aged ; Diagnosis, Differential ; Female ; Humans ; Male ; Middle Aged ; Nasal Cavity ; microbiology ; pathology ; Nasal Mucosa ; microbiology ; pathology ; Nose Diseases ; microbiology ; pathology ; Palatine Tonsil ; microbiology ; pathology ; Pharyngeal Diseases ; microbiology ; pathology ; Syphilis ; microbiology ; pathology ; Syphilis Serodiagnosis ; Treponema pallidum ; isolation & purification ; Young Adult

OBJECTIVETo establish a multiplex polymerase chain reaction (M-PCR) assay for simultaneous detection of pathogens causing genital ulcer disease (GUD).

METHODSBased on the gene-specific region of the following pathogens: Chlamydia trachomatis omp1/ompb, herpes simplex virus (HSV) DNA polymerase, Treponema pallidum tpp47, Haemophilus ducreyi 16s rRNA, four sets of primers were designed and an M-PCR assay was developed to detect four pathogens in one test. The assay was evaluated with diagnostic result of golden standard for each pathogen.

RESULTSOf the 51 clinical samples, M-PCR showed slightly higher positive rate (47.1%) of HSV than cell culture (23.6%). Meanwhile, the positive rate of T. pallidum detected by M-PCR and dark-field microscopy was 19.6% (10/51) and 15.7% (8/51), respectively. Only one sample was positive for H. ducreyi and no sample was positive for C. trachomatis detected by both M-PCR assay and culture.

CONCLUSIONThis primary study indicated that M-PCR assay can simultaneously and rapidly detect the four etiologic pathogens causing GUD.

Chlamydia trachomatis ; genetics ; isolation & purification ; DNA Primers ; Haemophilus ducreyi ; genetics ; isolation & purification ; Herpesvirus 1, Human ; genetics ; isolation & purification ; Herpesvirus 2, Human ; genetics ; isolation & purification ; Humans ; Polymerase Chain Reaction ; methods ; Sexually Transmitted Diseases ; complications ; microbiology ; virology ; Treponema pallidum ; genetics ; isolation & purification ; Ulcer ; complications ; microbiology ; virology