Using the Delphi method to establish nursing-sensitive quality indicators for ICU nursing in China.

In this study, the Delphi method was used to develop evidence-based indicators of intensive care unit (ICU) nursing quality of care in China. Nursing quality indicators reflect elements of patient care that are directly affected by nursing practice. A comprehensive literature search identified 2,857 potentially relevant articles. From the 50 articles that were included in this study, researchers identified 38 commonly used nursing quality indicators. A panel of experts reduced these to 20, which were then subjected to two rounds of Delphi discussion by a different panel, and a final consensus was achieved. The 20 indicators were grouped into three dimensions: structure, process, and outcome (including adverse consequences). The agreement among the experts for the 20 indicators was high. These evidence-based nursing quality indicators provide for ease in data collection and a basis for clinical application and improvement in the quality of ICU nursing throughout China.

[Analysis of Antibodies with Superior Antigen-recognition in Serum from Patients Infected by Necator americanus].

Objective: To evaluate the reactivity of adult hookworm antigens to serum from patients with hookworm disease, and analyze in the serum class- or subclass-specific antibodies that show superior antigen recognition. Methods: Sera from healthy participants, patients infected by Necator americanus and those with other parasitic infections were processed for ELISA, which used raw antigens extracted from adult worms of Necator americanus as the coating antigen, and different classes or subclasses of anti-human antibody labeled with HRP as the secondary antibody. The sensitivity and specificity of the assay with various secondary antibodies were compared. Results: The ELISA using IgM, IgD，IgE, IgG, IgG1, IgG2, IgG3, or IgG4 as the secondary antibody showed a sensitivity of 41.84%, 2.04%, 1.02%, 92.93%, 19.39%, 25.51%, 17.35%, and 88.78%, respectively; specificity of 77.61% 97.01%, 92.54%, 79.10%, 95.52%, 92.53%, 92.53%, and 92.53%, respectively; and diagnostic efficiency of 56.36%, 40.61%, 38.18%, 87.88%, 50.30%, 52.7%, 47.88%, and 90.30%, respectively. The sensitivity when using IgG4 and IgG as the secondary antibody had far exceeded that when using IgM, IgD, IgE, and other three subclasses of IgG (P<0.05). There was no difference in sensitivity between tests using IgG4 and IgG (χ2=1.61, P＞0.05). However, the test using IgG4 revealed significantly higher specificity than that using IgG (χ2=4.97, P＜0.05). Conclusion: Use of IgG4 as the enzyme-linked secondary antibody shows advantages in overall diagnostic efficiency over other classes/subclasses in ELISA.

[Diagnostic potential of five natural antigens from Echinococcus granulosus in the patients of cystic echinococcosis with different clinical status].

OBJECTIVE: To analyze the characteristics of serum antibody reactivity of cystic echinococcosis (CE) patients with different clinical status towards five native antigens obtained from Echinococcus granulosus (Eg). METHODS: The protoscolex somatic soluble antigen (EgPS), crude hydatid cyst fluid antigen (EgHF), partially purified hydatid fluid antigen (Burstein's antigen, EgBu), adult somatic soluble antigen (EgAs) and the native antigen B (EgAgB) were pre- pared. 369 serum samples from CE patients and 281 sera samples from healthy individuals were examined for the antibodies against 5 native antigens with indirect ELISA. The serologic results were classified according to clinical status, and the statistical analyses were carried out to understand the relationship between the results of different antigen-ELISA and the clinical status of patients. RESULTS: The results of EgBu, EgAS and EgAgB-ELISA showed that the antibody positive rate in hepatic CE patients [74.1% (212/286), 73.4% (210/286), 63.6% (182/286)] was significantly higher than that of other groups (including renal CE and pelvic CE, 1/8, 2/8, 1/8) (P < 0.05). Except EgAS, the S/N value of other groups examined by the rest four antigen-ELISA (EgPS: 3.10, EgHF: 2.40, EgBu: 1.60, EgAgB: 2.38) was also significantly lower than that of hepatic CE patients (3.73, 3.65, 4.40, and 3.61) (P < 0.05). EgBu, EgAS and EgAgB-ELISA results showed that the antibody positive rate in sera of recurrent CE patients [82.4% (150/182), 86.3% (157/182), 70.9% (129/182)] and the S/N value (5.54, 3.23, 3.75) were significantly higher than that of primary patients [positive rate: 67.4% (126/187); 63.6% (119/187); 57.2% (107/187); S/N value: 4.20, 2.70, 3.75] (P < 0.05). The S/N value detected by EgPS-ELISA and the positive rate examined by EgAgB-ELISA significantly increased with the increasing of the number of operations (P < 0.05), reached 4.23 and 91.7% (11/12), respectively, in the patients with > or = 4 times of operations. The positive rate and S/N value of EgAS-ELISA and EgAgB-ELISA increased with the number of hydatid cysts in patients (P < 0.05), reached 90.5% (19/21), 76.2% (16/21), and 3.97, 4.42, respectively, in patients with at least 4 cysts. Among the five antigen-ELISA, the positive rate increased with the cyst diameter (P > 0.05). The S/N value of EgHF-ELISA and EgAS-ELISA increased significantly with the cyst diameter (P < 0.05), reached 3.66 and 3.69, respectively, when the cyst diameter was > or = 15.1 cm. ROC analysis result showed that among the 5 native antigen-ELISA, the AUC(ROC) was highest in patients with cysts at CE2 stage (EgPS: 0.988 +/- 0.009, EgHF: 0.957 +/- 0.013, EgBu: 0.969 +/- 0.011, EgAs: 0.910 +/- 0.024, EgAgB: 0.894 +/- 0.021), EgAgB-ELISA presented the lowest AUC(RCO) of 0.267 +/- 0.031 in patients with cysts at CE5 stage. Except EgAgB, the positive rate of another 4 antigen-ELISA in detection of patients with cysts at CE 2 stage [EgPS: 97.2% (69/71), EgHF: 93.0% (66/71), EgBu: 88.7% (63/71), EgAs: 85.9% (61/71)] was slightly higher than that of the patients with cysts at CE1 stage, and then promptly reduced in patients with cysts at CE5 stage (EgPS: 56.3%, EgHF: 43.8%, EgBu: 12.5%, EgAs: 12.5%). In the patients with cysts at CE5 stage, the S/N value of the five antigen-ELISA was lowest (EgPS: 2.29, EgHF: 1.50, EgBu: 1.11, EgAs: 0.78, and EgAgB: 1.11). CONCLUSION: Compared with the other three antigens, the EgPS and EgAgB antigens have higher antigenicity, sensitivity, and specificity. The sera of hepatic CE patients are more reactive to the five native antigens than the other clinical types.

Differential diagnosis of cystic and alveolar echinococcosis using an immunochromatographic test based on the detection of specific antibodies.

Human cystic and alveolar echinococcoses are zoonotic diseases caused by the larval stages of Echinococcus granulosus and Echinococcus multilocularis, respectively. As the diseases are co-endemic in many areas of the world, a simple and rapid test for the differential diagnosis of cystic echinococcosis (CE) and alveolar echinocoocosis (AE) is needed. Here, we describe the development of an immunochromatographic test (ICT) using crude hydatid cyst fluid and a recombinant 18-kDa protein (rEm18) as antigens for the detection of E. granulosus and E. multilocularis antibodies in serum samples. The ICT was evaluated with serum samples from 195 echinococcosis patients from different endemic areas in northwestern China. These included 144 from CE patients, 51 from AE patients, 67 from patients with other parasitic diseases, 13 from patients with serous hepatic cysts, and 60 from healthy individuals. The sensitivity and specificity of the ICT for CE were 91.0 and 96.9% and for AE were 98.0 and 99.3% with diagnostic efficiencies of 94.1 and 99.1%, respectively. No significant differences and high degrees of agreement were found between the ICT and an enzyme-linked immunosorbent assay for both CE and AE. Five serum samples from cysticercosis patients and one serum sample from a healthy control were found positive for CE with the ICT. These findings indicate that this test allows for discrimination between both forms of human echinococcosis. In conclusion, the ICT developed in this study is a promising tool for the simultaneous detection and discrimination of CE and AE. This test will be useful for serodiagnosis of CE and AE in clinical settings and screening programs.

[Establishment and evaluation of colloid gold labeled immunochromatographic strip test for rapid diagnosis of alveolar echinococcosis].

OBJECTIVE: To establish and evaluate a colloid gold immunochromatographic strip test for the diagnosis of alveolar echinococcosis. METHODS: Total RNA was prepared from Echinococcus multilocularis protoscoleces collected from Xinjiang Uygur Autonomous Region. Em18 gene was obtained by reverse transcription-polymerase chain reaction (RT-PCR). The PCR product was sequenced and cloned into pGEX-3X vector. The recombinant plasmid was expressed and induced by isopropyl-beta-D-thiogalactopyranoside (IPTG) to obtain recombinant protein. The anti-human IgG monoclonal antibodies was conjugated with colloid gold as detecting reagent; the recombinant Em18 antigen and goat anti-mouse IgG were immobilized on nitrocellulose in proper position. The prepared immunochromatographic strip was evaluated using serum samples from patients with alveolar echinococcosis (56), cystic echinococcosis (87), cysticercosis (30), schistosomiasis japonica (10), toxoplasmosis (10) and healthy subjects (50) . Comparison between the immunochromatographic strip test and ELISA was made by kappa statistics. RESULTS: Sensitivity detected by the immunochromatographic strip test was 92.9% (52/56). The cross-reactivity to cystic echinococcosis and cysticercosis was 9.2% (8/87) and 3.3% (1/30), respectively. There was no cross reactivity with schistosomiasis japonica and toxoplasmosis. 4 samples out of 50 healthy people showed false positive reaction. The overall specificity was 93.0 (174/187). Sensitivity and specificity both showed no statistical difference between immunochromatographic strip test and ELISA. High degree of agreement was observed between the strip test and ELISA (kappa = 0.98). CONCLUSION: The developed immunochromatographic strip test using recombinant Em18 antigen as coated antigen is a sensitive, specific, simple and rapid assay for diagnosing alveolar echinococcosis.

[Detection and species identification of two imported cases of cutaneous leishmaniasis].

OBJECTIVE: To diagnose and identify pathogen of two suspected cases of cutaneous leishmaniasis. METHODS: Two cases of dermatosis with several major ulcers on the skin were examined, who worked and returned from Algeria (case 1) and Saudi Arabia (case 2), respectively. The stained smears of skin tissue from lesions were observed by microscope. Extravasate from lesions was cultured in NNN medium to search protozoan parasites, which were obtained by centrifugation. Two pairs of species-specific primers, ITS1-ITS2 and K13A-K13B, were used to amplify inter-nal transcribed spacer of rDNA and kinetoplast DNA, respectively. The products were sequenced and analyzed by Blast. RESULTS: There were Leishmania amastigotes in the tissue smear of case 2, while none in that of case 1. Promastigotes were found in culture medium of both cases. The PCR products of ITS1-ITS2 and K13A-K13B from 2 cases were about 330 bp and 120 bp with respective homology of 100% and 96% to corresponding sequences of Leishmania major. The accession numbers of 4 sequences were JF831924-JF831927. CONCLUSION: Two cases of dermatosis are diagnosed as imported cutaneous leishmaniasis and the pathogen is L. major.

Label-Free Quantitative Proteomic Analysis of Three Strains of Viscerotropic Leishmania Isolated from Patients with Different Epidemiological Types of Visceral Leishmaniasis in China.

BACKGROUND: There are three epidemiological types of visceral leishmaniasis in China, which are caused by Leishmania strains belonging to the L. donovani complex. The mechanisms underlying their differences in the population affected, disease latency, and animal host, etc., remain unclear. We investigated the protein abundance differences among Leishmania strains isolated from three types of visceral leishmaniasis endemic areas in China. METHODS: Promastigotes of the three Leishmania strains were cultured to the log phase and harvested. The protein tryptic digests were analyzed with liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS), followed by label-free quantitative analysis. The MS experiment was performed on a Q Exactive mass spectrometer. Raw spectra were quantitatively analyzed with the MaxQuant software (ver 1.3.0.5) and matched with the reference database. Differentially expressed proteins were analyzed using the bioinformatics method. The MS analysis was repeated three times for each sample. RESULTS: A total of 5012 proteins were identified across the KS-2, JIASHI-5 and SC6 strains in at least 2 of the three samples replicate. Of them, 1758 were identified to be differentially expressed at least between 2 strains, including 349 with known names. These differentially expressed proteins with known names are involved in biological functions such as energy and lipid metabolic process, nucleotide acid metabolic process, amino acid metabolic process, response to stress, cell membrane/cytoskeleton, cell cycle and proliferation, biological adhesion and proteolysis, localization and transport, regulation of the biological process, and signal transduction. CONCLUSION: The differentially expressed proteins and their related biological functions may shed light on the pathogenicity of Leishmania and targets for the development of vaccines and medicines.

Continuous Infusion of Angiotensin IV Protects against Acute Myocardial Infarction via the Inhibition of Inflammation and Autophagy.

Acute myocardial infarction (AMI) is a major cause of morbidity and mortality worldwide. Angiotensin (Ang) IV possesses many biological properties that are not yet completely understood. Therefore, we investigated the function and mechanism of Ang IV in AMI in in vivo and in vitro conditions. AMI was performed by ligation of the left anterior descending coronary artery (LAD) in male C57 mice. Ang IV was continuously infused by a minipump 3 d before AMI for 33 d. The neonatal rat ventricular myocytes (NRVCs) were stimulated with Ang IV and cultured under hypoxic conditions. In vivo, Ang IV infusion significantly reduced the mortality after AMI. By the 7th day after AMI, compared with the AMI group, Ang IV reduced the inflammatory cytokine expression. Moreover, terminal deoxyribonucleotidyl transferase- (TDT-) mediated dUTP nick-end labeling (TUNEL) assay showed that Ang IV infusion reduced AMI-induced cardiomyocyte apoptosis. Compared with AMI, Ang IV reduced autophagosomes in cardiomyocytes and improved mitochondrial swelling and disarrangement, as assessed by transmission electron microscopy. By 30th day after AMI, Ang IV significantly reduced the ratio of heart weight to body weight. Echocardiography showed that Ang IV improved impaired cardiac function. Hematoxylin and eosin (H&E) and Masson staining showed that Ang IV infusion reduced the infarction size and myocardial fibrosis. In vitro, dihydroethidium (DHE) staining and comet assay showed that, compared with the hypoxia group, Ang IV reduced oxidative stress and DNA damage. Enzyme-linked immunosorbent assay (ELISA) showed that Ang IV reduced hypoxia-induced secretion of the tumor necrosis factor- (TNF-) ɑ and interleukin- (IL-) 1ß. In addition, compared with the hypoxia group, Ang IV reduced the transformation of light chain 3- (LC3-) I to LC3-II but increased p62 expression and decreased cardiomyocyte apoptosis. Overall, the present study showed that Ang IV reduced the inflammatory response, autophagy, and fibrosis after AMI, leading to reduced infarction size and improved cardiac function. Therefore, administration of Ang IV may be a feasible strategy for the treatment of AMI.

[Decomposition dynamics and driving factors of manure in reclaimed soils in coal mining area]. / ç ¤çŸ¿åŒºå¤åž¦åœŸå£¤ä¸­æœ‰æœºè‚¥çš„åˆ†è§£åŠ¨æ€åŠå ¶é©±åŠ¨å› ç´ .

Understanding the decomposition dynamics and driving factors of manure in the soil subjected to different reclaimed years could provide theoretical basis to rational utilization of manure and soil fertility improvement in coal mining area. Cattle manure and pig manure were mixed with soils subjected to different reclaimed years (one year, R1; 10 years, R10; and 30 years, R30) at the ratio of manure carbon to soil mass of 4 to 100, so as to examine manure decomposition characteristics using the nylon mesh bag (15 cm deep of soil buried) in the Shanxi coal mine reclamation area, with no manure addition as control (CK). Soil samples were collected at day 12, 23, 55, 218, 281, and 365 to measure the contents of soil manure residual, soil microbial biomass carbon (MBC), and dissolved organic carbon (DOC). The contributions of soil properties, manure properties, and hydrothermal condition to manure decomposition were quantified. The results showed that the decomposition rates of pig manure were significantly higher than cattle manure. The humification coefficient of pig manure (average 46.3%) was lower than that of cattle manure (average 71.7%). The humification coefficient of pig manure was significantly lower in the 30-year reclaimed soil (44.5%) compared to the 1-year and 10-year reclaimed soil (average 47.2%). There was no significant difference in the humification coefficient of cattle manure among the three reclaimed soils. The proportion and decomposition rate constant of labile carbon pool of pig manure and cattle manure were significantly different, with values of 52% and 26%, and 0.00085 and 0.00074 ℃-1, respectively. The positive effect of pig manure on MBC and DOC in reclaimed soil was significantly higher than that of cattle manure over 0-218 days, but no difference over 281-365 days. The magnitude of the enhancement of MBC and DOC in those three reclaimed soils after manure amendments showed a similar trend of R1 >R10 ≈ R30. Results of variance partitioning analysis showed that manure decomposition was mainly controlled by manure properties (17.9%) when considering soil properties, manure properties, and hydrothermal condition. In conclusion, the decomposition of pig manure but not cattle manure was regulated by reclamation year. Cattle manure, with higher humification coefficient than pig manure, was recommended for reclaimed mining area to improve soil fertility.

Is Dynesys dynamic stabilization system superior to posterior lumbar fusion in the treatment of lumbar degenerative diseases?

Dynesys, a pedicle-based dynamic stabilization system, was introduced to overcome some undesirable complications of fusion procedures. Nevertheless, the theoretical advantages of Dynesys over fusion have not been clearly confirmed. The purpose of this editorial was to compare clinical and radiological outcomes of patients who underwent Dynesys system with those who underwent posterior lumbar fusion according to the existing literature and to see if the application of the Dynesys system is superior to the traditional lumbar fusion surgery. According to published clinical reports, the short-term effects of the Dynesys dynamic stabilization system are similar to that of traditional lumbar fusion surgery. Three comparative studies of Dynesys dynamic stabilization and fusion surgery with medium-term follow-up are encouraging. However, the results from four single-treatment-arm and small-sample studies of case series with long-term follow-up were not encouraging. In the present circumstances, it is not possible to conclude that the Dynesys dynamic stabilization system is superior to fusion surgery for lumbar degenerative diseases.

Contrast in soil microbial metabolic functional diversity to fertilization and crop rotation under rhizosphere and non-rhizosphere in the coal gangue landfill reclamation area of Loess Hills.

Very poor reclaimed soil quality and weak microbial activity occur in the reclamation area of a coal gangue landfill in the Loess Hills. The fourth and fifth years after farmland soil was reclaimed were studied, and the changes in and carbon source utilization characteristics of rhizosphere (R) and non-rhizosphere (S) soil microorganisms under organic and inorganic (OF), inorganic (F), and organic (O) fertilizer application and a control treatment (CK) in soybean (S) and maize (M) rotation systems were compared and analysed in Guljiao Tunlan, Shanxi Province, China. Biolog-EcoPlate technology was used to analyse the mechanism of soil characteristic change from the perspective of soil microbial metabolism function to provide a theoretical basis for reclamation and ecological reconstruction in this area. The average well colour development (AWCD) absorption and Shannon-Wiener index values of soybean and maize rhizosphere microorganisms were higher than those of non-rhizosphere microorganisms, and the mean value of the fertilizer treatment was higher than that for CK. Principal component analysis shows the main carbon sources that impact the functional diversity of the soybean rhizosphere and non-rhizosphere soil communities are a-cyclodextrin, a-D-lactose, ß-methyl D-glucoside, and glucose-1-phosphate and L-phenylalanine, while those for the maize rhizosphere and non-rhizosphere soil communities are D-cellobiose, glucose-1-phosphate, ß-methyl D-glucoside, methyl pyruvate, D-galactosate gamma lactone, D-mannitol, N-acetyl-D-glucosamine, D-galactosalonic acid, and L-serine. The comprehensive utilization score of the non-rhizosphere soil carbon source in the maize season increased with respect to that in the soybean season, and the maximum increase was 1.09 under the OF treatment. Redundancy analysis showed that the soil nutrient factors driving the changes in the metabolic function diversity index values of the rhizosphere and non-rhizosphere soil microbial communities in the different crop seasons in the reclamation area differed, but they were all related to the soil organic matter and available phosphorus. This may explain why OF treatment is the most beneficial to soil fertility under the rotation system in mining reclamation areas.

Contributions of the National Institute of Parasitic Diseases to the control of visceral leishmaniasis in China.

Visceral leishmaniasis (VL) caused by Leishmania spp. is an important vector-borne disease prevalent in China. VL was rampant in the vast area of China north of the Yangtze River before the founding of the People's Republic of China in 1949. As a result of strenuous interventions, the disease was basically eliminated in most of the former epidemic areas in 1958-60. At present, only sporadic cases occur in the western regions of China. In the process, National Institute of Parasitic Diseases at China CDC and the Chinese Center for Tropical Diseases Research (NIPD-CTDR) have achieved great impact in controlling the diseases as well as in research on Leishmania spp. This review summarized the contribution of experts from NIPD-CTDR to the control and elimination of VL in various aspects, such as understanding the epidemiological features of VL, confirmation of VL vectors and their distribution, development of control tools including diagnostics and insecticides, monitoring and evaluation supported by information management, technical supports to the control programmes, as well as analysis of the challenges faced. At the same time, it puts forward constructive suggestions for the ultimate interruption of VL transmission in China.

Contribution to the echinococcosis control programme in China by NIPD-CTDR.

As a zoonotic parasitosis caused by the parasitism of Echinococcus larvae, echinococcosis imposes serious disease and economic burdens on human beings and society, and is thus a global public health issue. Its complex life history, wide distribution, the combined influence of various epidemic factors, coupled with the unique natural environment, customs, and religious beliefs in endemic areas, pose a huge challenge to the national echinococcosis control programme in China. Accurate early detection and confirmation of diagnosis of echinococcosis, the use of effective drugs, real-time surveillance of the infection status of populations and various hosts, controlling the source of infection, and blocking the route of transmission are of enormous significance for control. In this paper, the work by NIPD-CTDR on the prevention and control of echinococcosis in China is reviewed, with a view to providing reference for the further promotion of the national echinococcosis control programme.

[A special type of discogenic low back pain: normal signal intensity of magnetic resonance imaging with abnormal discography].

OBJECTIVE: To report a group of special patients showing a normal signal intensity of MRI with a positive discography and discuss its pathogenesis and clinical significance. METHODS: From August 2003 to November 2008, 288 patients with chronic low back pain were treated. Of these patients, 12 showed a normal intensity signal of MRI in lumbar intervertebral discs with a positive discography. There were 7 males and 5 females aged 20 - 44 years with an average of 29.6 years. The duration of disease was 8 months to 3 years with an average of 1.8 years. Dallas CT grading method was used for assessing the degree of annular disruption. RESULTS: Of these 12 patients, 33 lumbar discs underwent discography. Twelve discs in 12 patients showed annular disruption and low back pain reproduction. Among 12 painful discs, 3 showed grade 2 annular disruption and 9 showed grade 3 annular disruption. CONCLUSION: Patients who are refractory to conservative care and have normal signal intensity in lumbar disc MRI and who are suggested as discogenic origin and might need further treatment such as minimal invasive surgery or lumbar interbody fusion should still be examined by discography.

[Field evaluation of gold-immunochromatographic assay for diagnosis of vivax malaria].

OBJECTIVE: To evaluate a gold-immunochromatographic assay (GICA) for malaria diagnosis in an endemic area of vivax malaria. METHODS: Blood samples were collected from febrile patients in 5 township-hospitals of Mengcheng County, Anhui Province, between September and October 2008. The samples were examined by GICA and microscopy under double blind condition and the results were compared. RESULTS: Among 292 blood samples, 181 were found P. vivax-positive by microscopy, and 163 were positive by GICA. Altogether, the coincidence of the two methods stood for 92.8% (271/292), including 108 negatives and 163 positives. 21 samples with discrepancy covered 18 microscopy positive but GICA negative, 3 microscopy negative but GICA positive. The GICA positive rate in patients with a parasitaemia of > 1,000 parasites/microl, 100-1,000 parasites/microl, and < 100 parasites/microl was 93.5% (115/123), 86.0% (43/50), and 62.5% (5/8), respectively. CONCLUSION: GICA is a useful diagnosis method for endemic area of vivax malaria.

Field evaluation of an immunochromatographic test for diagnosis of cystic and alveolar echinococcosis.

BACKGROUND: The larval stages of the tapeworms Echinocoocus granulosus and Echinococcus multilocularis are the causative agents of human cystic echinococcosis (CE) and human alveolar echinococcosis (AE), respectively. Both CE and AE are chronic diseases characterised by long asymptomatic periods of many years. However, early diagnosis of the disease is important if treatment and management of echinococcosis patients are to be successful. METHODS: A previously developed rapid diagnostic test (RDT) for the differential detection of CE and AE was evaluated under field conditions with finger prick blood samples taken from 1502 people living in the Ganzi Tibetan Autonomous Prefecture, China, a region with a high prevalence for both forms of human echinococcosis. The results were compared with simultaneously obtained abdominal ultrasonographic scans of the individuals. RESULTS: Using the ultrasonography as the gold standard, sensitivity and specificity, and the diagnostic accuracy of the RDT were determined to be greater than 94% for both CE and AE. For CE cases, high detection rates (95.6-98.8%) were found with patients having active cysts while lower detection rates (40.0-68.8%) were obtained with patients having transient or inactive cysts. In contrast, detection rates in AE patients were independent of the lesion type. The positive likelihood ratio of the RDT for CE and AE was greater than 20 and thus fairly high, indicating that a patient with a positive test result has a high probability of having echinococcosis. CONCLUSIONS: The results suggest that our previously developed RDT is suitable as a screening tool for the early detection of human echinococcosis in endemic areas.

[Establishment and evaluation of colloid gold labeled immunochromatographic strip test for rapid diagnosis of malaria].

OBJECTIVE: To establish and evaluate a gold immunochromatographic strip test for detection and differentiation of Plasmodium vivax and P. falciparum. METHODS: The monoclonal antibodies, F4H12, G4C9 and D8F7, were conjugated with colloid gold as detecting reagent; monoclonal antibody B2G10 (against P. vivax/ P. falciparum) and D6A7 (only against P. falciparum) were immobilized on nitrocellulose in proper position. Blood samples from 107 febrile patients from endemic area of malaria and 17 patients with visceral leishmaniasis were used for evaluating the specificity. Blood samples of malaria patients (110 with P. vivax and 54 with P. falciparum) were used for evaluating the sensitivity. RESULTS: 5 samples out of 107 febrile patients and 17 patients with visceral leishmaniasis showed false positive reaction with a specificity of 96.0% (119/124), all the 17 samples from patients with visceral leishmaniasis were negative. 164 blood samples of malaria patients showed a sensitivity of 92.3% (153/164), 92.7% (102/110)and 94.4% (51/ 54) for patients infected with P. vivax or P. falciparum, respectively. CONCLUSION: The immunochromatographic strip test based on antigen-capturing is a sensitive, specific, simple and rapid assay for malaria diagnosis.

[Asymptomatic Leishmania infection in human population of Wenxian County, Gansu Province].

OBJECTIVE: To analyze the status of Leishmania infantum asymptomatic infection in human population of a Kala-azar endemic area in Wenxian County, Gansu Province, and to evaluate the tests used. METHODS: Blood samples were tested by PCR using two pairs of primers, RV1-RV2 and K13A-K13B, for detecting Leishmania-specific DNA. ELISA and rK39-dipstick were used to detect Leishmania-specific antibodies. RESULTS: The positive rate of PCR, ELISA and rK39-dipstick was 30.9%(83/269). 24.2%(65/269) and 0 (0/269) respectively. CONCLUSION: The prevalence of asymptomatic infection of L. infantum in humans is high in the area. PCR test based on RV1-RV2 and K13A-K13B primer pairs is a sensitive and specific method for detecting the asymptomatic infection.

[Study on PCR method for detecting the asymptomatic infection of Leishmania infantum].

OBJECTIVE: To establish PCR method for the detection of the asymptomatic infection of Leishmania infantum. METHODS: Six primer pairs were selected for detecting Chinese strain of L. infantum by optimizing conditions which affect amplification. Their sensitivity and specificity were compared by using DNAs extracted from human blood seeded with cultured L. infantum promastigotes (MHOM/CN/86/GS) as template. Blood samples of the inhabitants without symptoms of visceral leishmaniasis in the endemic area were analyzed with two selected primer pairs with good sensitivity and specificity. RESULTS: The specificity of all six primer pairs reached 100%, and the sensitivity varied among the primer pairs. The primer pairs RV1-RV2 (0.1 parasite/ml blood) and K13A-K13B (1 parasite/ml blood) were most sensitive. Leishmania DNA was detected in 33% (33/100) and 30% (30/100) human blood samples by RV1-RV2 and K13A-K13B primer pairs respectively. CONCLUSION: This study suggests that RV1-RV2 and K13A-K13B primer pairs are suitable in detecting the asymptomatic infection of L. infantum, and the prevalence of the asymptomatic infection is high in human population in the endemic area.

Development of an Immunochromatographic Test for Diagnosis of Visceral Leishmaniasis Based on Detection of a Circulating Antigen.

BACKGROUND: Visceral leishmaniasis (VL) is a life-threatening disease caused by protozoan parasites of the Leishmania donovani complex. Early case detection followed by adequate treatment is essential to the control of VL. However, the available diagnostic tests are either invasive and require considerable expertise (parasitological demonstration of the parasite in tissue smears) or unable to distinguish between past and active infection (serological methods). Therefore, we aimed to develop a lateral flow assay in the form of an immunochromatographic test (ICT) device based on the detection of a circulating Leishmania antigen using monoclonal antibodies (mAbs). METHODOLOGY/PRINCIPAL FINDINGS: mAbs were produced by fusion of murine myeloma cells with splenocytes isolated from a mouse immunized with L. donovani soluble crude antigen. Out of 12 cloned hybridoma cell lines, two secreted mAbs recognizing the same leishmanial protein. These mAbs were used to produce an ICT as a sandwich assay for the detection of circulating antigen in serum and blood samples. The ICT was evaluated with 213 serum samples from VL patients living in VL endemic areas in China, and with 156 serum samples from patients with other diseases as well as 78 serum samples from healthy donors. Sensitivity, specificity and diagnostic efficiency of the new ICT was 95.8%, 98.7% and 97.3%, respectively. Compared with a commercially available antibody detecting ICT, our antigen-based ICT performed slightly better. CONCLUSION/SIGNIFICANCE: The newly developed ICT is an easy to use and more accurate diagnostic tool which fulfils the performance and operational characteristics required for VL case detection under field and laboratory conditions. As our ICT detects a circulating antigen, it will also be useful in monitoring treatment success and diagnosing VL in immunocompromised patients.