Pyosalpinx: not always a sexual transmitted disease? Pyosalpinx caused by Plesiomonas shigelloides in an immunocompetent host

Plesiomonas shigelloides are ubiquitous Gram-negative bacteria that are found in fresh or marine water, particularly in tropical or warm climates; they were recently implicated in diarrhoeal disease. Patients usually present with a history of recent travel to tropical regions or consumption of uncooked seafood. Extraintestinal disease has rarely been reported, occurring generally in neonates or immunocompromised patients, and is often fatal. We report a case of right pyosalpinx due to P. shigelloides acquired by swimming in contaminated water. Laparoscopic salpingectomy led to a good outcome.     

医学图像计算机辅助诊断数据平台研究

缺乏统一的数据存储模型及相关研究工具,是制约计算机辅助诊断研究中数据、算法等研究成果共享的最主要因素。针对计算机辅助诊断协同研究中的这一问题,提出并构建了用于医学图像计算机辅助诊断协同研究的数据平台。首先对计算机辅助诊断协同研究中的影像数据、过程数据以及“金标准”数据需求进行了分析,提出了统一的数据模型用于数据存储和表示,采用Oracle数据库实现;分析了病例获取、“金标准”标注、分割、特征提取以及算法评估等研究阶段的业务需求,设计并完成了相应的研究工具;最后基于DCMTK采用VC++编程实现了数据平台。数据平台应用于肺癌和脑胶质瘤辅助诊断研究,较好地解决了医学图像计算机辅助诊断研究过程中的数据存储、病例获取、标注、研究结果存储及评估问题,达到了共享研究成果的目的,对提高计算机辅助诊断研究有很好的促进作用。     

Diagnosis of Visceral Leishmaniasis by Enzyme-Linked Immunosorbent Assay Using Urine Samples

A diagnostic method has been developed to detect anti-Leishmania donovani immunoglobulin G (IgG) in urine by enzyme-linked immunosorbent assay (ELISA). In measuring anti-L. donovani IgG, IgA, and IgM in urine, the method performed best in the detection of IgG. The sensitivity and specificity of the assay were determined with panels of urine samples from 62 visceral leishmaniasis (VL) patients, 59 healthy controls from areas of endemicity, 53 healthy controls from areas of nonendemicity, 59 malaria patients, 13 tuberculosis patients, 23 cutaneous leishmaniasis patients, and 7 patients with other diseases. Using L. donovani promastigote crude antigen, the test had 93.5% sensitivity (58 positives of 62 VL patient samples) and 89.3% specificity (191 negatives of 214 non-VL patient samples). The ELISA with acetone-treated L. donovani promastigote antigen raised the sensitivity and specificity to 95.0 and 95.3%, respectively. Western blot analysis revealed that most of the samples that cross-reacted with crude antigen in ELISA did not recognize any antigenic component of L. donovani crude antigen. We also checked 40 serum samples from the same group of VL patients for anti-L. donovani IgG and got 90.0% sensitivity with both crude and acetone-treated antigens. As collection of urine is much easier than collection of serum, the detection of anti-L. donovani IgG in urine with acetone-treated antigen will be useful in epidemiological studies. It could be an adjunct of laboratory diagnosis.     

Diagnosis of Trichomonas vaginalis Infection by PCR Using Vaginal Swab Samples

Trichomonas vaginalis infection is the most prevalent nonviral sexually transmitted disease (STD) in the world. A PCR test using vaginal swab samples for the detection of T. vaginalis was developed to add T. vaginalis infection to the growing list of STDs that can be detected by DNA amplification techniques. A primer set, BTUB 9/2, was designed to target a well-conserved region in the beta-tubulin genes of T. vaginalis. All strains (15 of 15) of T. vaginalis tested were successfully detected by PCR giving a single predicted product of 112 bp in gel electrophoresis. No such targeted product was amplified with DNA from Trichomonas tenax, Trichomonas gallinae, Chlamydia trachomatis, Neisseria gonorrhoeae, Giardia lamblia, Chilomastix sulcatus, Dientamoeba fragilis, and Entamoeba histolytica. An optimal analytical sensitivity of one T. vaginalis organism per PCR was achieved. Culture, performed with the Inpouch TV culture system, was examined daily with a light microscope to identify T. vaginalis. Twenty-three of 350 (6.6%) vaginal swab samples from women attending an army medical clinic were culture positive for T. vaginalis. Of these culture positive specimens, PCR detected 22 of 23 (96%) with primer set BTUB 9/2, and wet preparation detected only 12 of 23 (52%). Seventeen specimens were BTUB 9/2-PCR positive and culture negative. Ten of these discordant specimens were determined to be as true positive by PCR using primer sets TVA 5-1/6 and/or AP65 A/B, which target different regions in the T. vaginalis genome, and seven were determined to be false positive. The sensitivity of BTUB 9/2-PCR was 97% and the specificity was 98%. The sensitivities of culture and wet preparation were 70 and 36%, respectively. The diagnosis of T. vaginalis infection by PCR is a sensitive and specific method that could be incorporated into a joint strategy for the screening of multiple STDs by using molecular amplification methods.     

Rapid Diagnosis of Azole-Resistant Aspergillosis by Direct PCR Using Tissue Specimens

We report the use of PCR techniques on a formalin-fixed and paraffin-embedded tissue specimen for direct detection of one dominant azole resistance mechanism in a case of disseminated invasive aspergillosis. Rapid detection of mutations associated with azole resistance directly in tissue significantly reduces diagnostic delay.Invasive infections due to Aspergillus fumigatus are associated with significant morbidity and mortality, although the prognosis of patients with invasive aspergillosis has improved with the clinical use of mold-active antifungal azoles, most notably voriconazole (9, 11). However, the survival of patients may be threatened by the emergence of azole resistance of aspergilli (1, 7, 13). Resistance is commonly due to point mutations in the cyp51A gene, which is the target for antifungal azoles (1, 4, 8, 13, 14). The isolates commonly exhibit a cross-resistant phenotype (4), and patients with azole-resistant disease may fail azole therapy (1, 7, 10, 12). One problem in the management of azole-resistant aspergillosis is the early detection of resistance as cultures are negative in up to 50% of patients with focal pulmonary lesions (2), and in vitro susceptibility testing takes at least 5 to 7 days to complete. In this report, molecular tools were utilized to rapidly confirm the diagnosis of disseminated azole-resistant aspergillosis.     

Diagnosis of Congenital Toxoplasmosis by Using a Whole-Blood Gamma Interferon Release Assay

Congenital toxoplasmosis in newborns is generally subclinical, but infected infants are at risk of developing ocular lesions. Diagnosis at birth relies mainly on serological tests. Cell-mediated immunity plays the major role in resistance to infection but is not routinely investigated for diagnostic purposes. Here, we describe a simple test based on the gamma interferon (IFN-γ) response after stimulation of whole blood by crude parasitic antigens. One milliliter of heparinized blood was centrifuged; plasma was kept for routine serological tests, and pellets were resuspended in culture medium. After 24 h of culture in the presence of crude Toxoplasma gondii antigen, the cells were centrifuged and the supernatant was assayed for IFN-γ. For 62 infants under 1 year of age born to mothers who were infected during pregnancy, the sensitivity and specificity of the test were 94% (with positive results for 16 of 17 infected infants) and 98% (with negative results for 44 of 45 uninfected infants), respectively. The false-negative result was for a treated baby who gave positive results after the withdrawal of treatment. The false positive was obtained for a 3-month-old baby. For a cohort of 124 congenitally infected patients between 1 and 30 years of age, the sensitivity of the assay was 100%. We present a simple test based on IFN-γ secretion to assess cell-mediated immunity in toxoplasmosis. As only 1 ml of blood is required to investigate humoral and cellular immunity, our assay is well adapted for the study of congenital toxoplasmosis in infants. Using purified antigens or recombinant peptides may improve the test performance.Toxoplasma gondii, a ubiquitous intracellular protozoan parasite, is an important cause of morbidity and mortality in congenitally infected individuals. Maternal infection may have serious consequences for the fetus (10). In other cases, infected newborns appear to be totally asymptomatic at birth but are at risk of developing retinal diseases during childhood or adolescence (26). For these patients, the diagnosis of the disease relies mainly on the detection of specific antibodies. Toxoplasma-specific immunoglobulin M (IgM) and IgA, which do not cross the placenta, are considered to be good markers of congenital infection. However, gestational age at maternal infection affects test performance (25), and at birth, the tests cannot detect more than 75% of infected babies (20). Because Toxoplasma-specific IgG crosses the placenta, its presence in the blood of newborns cannot be considered a marker of congenital infection. Maternally transferred IgG usually disappears within 6 to 12 months (20). Therefore, uninfected infants born to mothers who seroconverted during pregnancy have to undergo regular sampling for serological testing for 1 year before congenital toxoplasmosis can be ruled out (16). Clinicians are seeking valid indicators of congenital infection to improve clinical decision making. T. gondii infection results in long-lasting cell-mediated immunity which is highly dependent upon the effector activity of T lymphocytes that produce gamma interferon (IFN-γ) (6). Surprisingly, few studies have investigated the potential role of cell immunity in diagnosis of the disease. Data in the literature are contradictory. An absence of stimulation of lymphocytes by T. gondii antigen in congenitally infected children has been reported previously (18, 27). Recently, Guglietta et al. (13), using synthetic peptides, detected age-related impairment of the specific T-cell response to parasitic antigen in congenital infections. Conversely, a recent publication reported that evaluation of T-cell immunity is important for an early and accurate diagnosis of congenital toxoplasmosis (3). By detecting CD25 expression by flow cytometry, we demonstrated previously that specific cell immunity is detectable in almost all infected patients, including newborns (11). In this study, we evaluate the performance of a whole-blood IFN-γ release assay for the diagnosis of congenital toxoplasmosis. In this clinical setting, the quantity of blood required is limited, and we therefore looked at the possibility of separating plasma from blood cells in order to conduct serological tests (the “gold standard”) and the IFN-γ assay with the same sample.     

Diagnosis of Mycobacterium microti Infections among Humans by Using Novel Genetic Markers

As a result of DNA typing of Mycobacterium microti isolates from animals in the United Kingdom and The Netherlands, we diagnosed four human M. microti infections. These are the first M. microti infections among humans to be reported. Three of the patients were immunocompromised and suffered from generalized forms of tuberculosis. The fourth patient was a 34-year-old immunocompetent male with a persistent cough and undefined X-ray abnormalities. Two of the M. microti infections were recognized by their IS6110 restriction fragment length polymorphism (RFLP) patterns, which showed a high degree of similarity with those of M. microti strains isolated from a pig and a ferret in The Netherlands. The two other human M. microti infections were recognized by using the recently developed DNA fingerprinting method, “spoligotyping,” directly on clinical material. All M. microti isolates from the United Kingdom and The Netherlands were found to contain an exceptionally short genomic direct repeat region, resulting in identical two-spacer sequence reactions in spoligotyping. In contrast, the highly similar IS6110 RFLP patterns of the vole strains from the United Kingdom differed considerably from the RFLPs of all M. microti strains isolated in The Netherlands, suggesting that geographic isolation led to divergent strains in the United Kingdom and on the continent.     

Diagnosis of Penicillium marneffei Infection by Quantitation of Urinary Antigen by Using an Enzyme Immunoassay

Penicillium marneffei is a major cause of opportunistic infection in patients with AIDS in north and northeastern Thailand. A method for the quantitation of P. marneffei antigen in urine was developed by using fluorescein isothiocyanate-labelled purified rabbit hyperimmune immunoglobulin G in an enzyme-linked immunosorbent assay. This method was evaluated with 33 patients with culture-proven penicilliosis and 300 controls (52 healthy subjects, 248 hospitalized patients without penicilliosis) from the same area in which penicilliosis is endemic. Urinary antigen was found in all 33 (100%) patients with penicilliosis, with a median titer of 1:20,480. With undiluted samples, 67 (27%) of 248 hospital patients and 3 (6%) of 52 healthy controls were reactive. At a cutoff titer of 1:40, the urine antigen detection assay had a diagnostic sensitivity of 97% and specificity of 98% (positive predictive value, 84%; negative predictive value, 99.7%). This test offers a valuable and rapid method for the diagnosis of penicilliosis in patients with AIDS and could be a useful addition to conventional diagnostic methods in areas in which penicilliosis is endemic.     

利用软组织力-位移测试系统诊断颈痛

通过局部软组织力-位移测试系统，对正常对照组、颈痛组的局部软组织力和位移定量检测；同时用目前诊断颈痛的方法作为对照，对局部软组织生物力学测试诊断颈痛的准确性、可靠性及临床作出评价，试图为颈部疼痛的诊断提供生物力学定性、定最指标。     

Rapid Diagnosis of Japanese Encephalitis by Using an Immunoglobulin M Dot Enzyme Immunoassay

Japanese encephalitis (JE) occurs in rural settings in southern and eastern Asia, where diagnostic facilities are limited. For the diagnosis of JE virus (JEV) infection, we developed a nitrocellulose membrane-based immunoglobulin M (IgM) capture dot enzyme immunoassay (MAC DOT) that is rapid, simple to use, requires no specialized equipment, and can distinguish JEV from dengue infection. In a prospective field study in southern Vietnam, 155 cerebrospinal fluid (CSF) and 341 serum samples were collected from 111 children and 83 adults with suspected encephalitis. The JEV MAC DOT, performed on site, was scored visually from negative to strongly positive by two observers, and the results were compared subsequently with those of the standard IgM capture enzyme-linked immunosorbent assay. For the 179 patients with adequate specimens, the MAC DOT correctly identified 59 of 60 JEV-positive patients and 118 of 119 JEV-negative patients (sensitivity [95% confidence intervals], 98.3% [92.1 to 99.9%]; specificity, 99.2% [95.9 to 100.0%]; positive predictive value, 0.98; negative predictive value, 0.99). The MAC DOT also correctly identified three patients with dengue encephalopathy. Admission specimens were positive for 73% of JE patients. Interobserver agreement for MAC DOT diagnosis was excellent (kappa = 0.94). The JEV MAC DOT is a simple and reliable rapid diagnostic test for JE in rural hospitals.     

Diagnosis of Babesiosis Using an Immunoblot Serologic Test

Although the current indirect immunofluorescent assay (IFA) diagnostic antibody test for human babesiosis is sensitive and specific, an immunoblot antibody test may be easier to standardize and to perform. Our objective, therefore, was to determine the efficacy of and develop interpretive criteria for an immunoblot antibody test for diagnosing acute human babesiosis using a Babesia microti whole-cell lysate as the antigen. We compared the reactivity of sera to a B. microti immunoblot assay in 24 human subjects experiencing symptoms and expressing laboratory evidence of babesiosis, 28 subjects who experienced Lyme disease, 12 subjects who experienced human granulocytic ehrlichiosis, and 51 subjects who reported no history of any of these diseases and whose sera did not react against B. microti antigen in an IFA test. Immunoblot strips were impregnated with proteins derived from the GI strain of B. microti that had been electrophoresed in an acrylamide sodium dodecyl sulfate gel, followed by electroblotting onto nitrocellulose membranes. The sera of all subjects who experienced babesiosis reacted against the B. microti antigen in the IFA and against at least one of nine immunoblot protein bands specific to B. microti. In contrast, none of the sera from people who appeared not to have experienced this infection reacted against the B. microti antigen in the IFA (compared to 4% in the immunoblot assay). When two reactive bands were considered as definitive, immunoblot test sensitivity was 96%, while specificity was 99% and predictive positivity and predictive negativity were 96 and 99%, respectively. Our B. microti immunoblot procedure shows promise as a sensitive, specific, and reproducible assay for routine clinical diagnosis of acute babesiosis.     

Development of Pathological Diagnosis Support System Using Micro-computed Tomography

In pathological diagnosis, the cutting position of pathological materials is subjectively determined by pathologists. This leads to a low cutting accuracy, which in turn may lead to incorrect diagnoses. In this study, we developed a system that supports the determination of the cutting position by visualizing and analyzing the internal structure of pathological material using micro-computed tomography (CT) before cutting. This system consists of a dedicated micro-CT and cutting support software. The micro-CT system has a fixture for fixing the target, enabling the scanning of easily deformable pathological materials. In the cutting support software, a function that interactively selects the extraction plane while displaying the volume rendering image and outputs a pseudo-histological image was implemented. Our results confirmed that the pseudo-histological image showed the fine structure inside the organ and that the latter image was highly consistent with the pathological image.     

联合超声技术诊断上肢深静脉血栓形成

目的探讨上肢深静脉血栓 (UEDVT) 的声像特征及超声诊断UEDVT的临床价值.方法对38例单侧上肢临床疑UEDVT的患者进行超声检查,包括二维加压超声、彩色血流多普勒显像(CFDI)、二维血流显像(B-flow)及脉冲多普勒技术(PWD)的综合使用.结果 26例经检查确诊UEDVT,其中4例合并有浅静脉血栓.11例正常,1例为胸廓出口综合征.完全栓塞42条,不完全栓塞21条.结论加压超声结合CDFI及/或B-flow对确定静脉血栓部位、静脉阻塞情况的判断有可靠的价值,为诊断UEDVT的首选方法,PWD对判断血流状态有帮助,但对判断血栓的存在不是必须的.     

联合超声技术诊断上肢深静脉血栓形成

目的　探讨上肢深静脉血栓 (UEDVT)的声像特征及超声诊断UEDVT的临床价值 .方法　对 3 8例单侧上肢临床疑UEDVT的患者进行超声检查 ,包括二维加压超声、彩色血流多普勒显像 (CFDI)、二维血流显像 (B -flow)及脉冲多普勒技术 (PWD)的综合使用 .结果　 2 6例经检查确诊UEDVT ,其中 4例合并有浅静脉血栓 . 11例正常 ,1例为胸廓出口综合征 .完全栓塞 42条 ,不完全栓塞 2 1条 .结论　加压超声结合CDFI及 /或B -flow对确定静脉血栓部位、静脉阻塞情况的判断有可靠的价值 ,为诊断UEDVT的首选方法 ,PWD对判断血流状态有帮助 ,但对判断血栓的存在不是必须的