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Journal, Genetic Engineering &... Dec 2018Leptospirosis is a widespread zoonotic disease caused by . Symptoms of disease range from mild symptoms to serious complications including, jaundice, pulmonary...
Leptospirosis is a widespread zoonotic disease caused by . Symptoms of disease range from mild symptoms to serious complications including, jaundice, pulmonary hemorrhage, renal and hepatic failure, which may prove fatal. Clinical presentations of this disease are similar with other febrile illness. Therefore, rapid and appropriated laboratory diagnostic tests are needed to aid clinical case identification. As these reasons, objective of this study is to develop and evaluate a simple latex agglutination test coating with recombinant leptospiral antigens, LipL32 for serodiagnosis of human leptospirosis. Firstly, gene was amplified from genomic DNA of serovar Pyrogenes. Then PCR product of gene was ligated with pGEX-2T plasmid, generating pGRK32 recombinant plasmid. Recombinant GST-LipL32 protein was overexpressed and subsequently purified by using Glutathione-Agarose Resin. Recombinant GST-Lipl32 protein was coated on latex beads for development latex agglutination test (LAT). The relative sensitivity, specificity and accuracy of the developed LAT were compared with indirect immunofluorescences assay (IFA) for detection of anti-leptospiral antibodies in 30 human leptospirosis samples, 30 healthy blood donor samples, 10 dengue fever positive samples, 10 scrub typhus positive samples, and 10 melioidosis samples. Results showed that the developed LAT showed sensitivity, specificity and accuracy: 66.66%, 86.66%, and 80.00%, respectively, comparing with IFA method. Moreover, Kappa analysis showed agreement rate of the two methods were 0.421. It concluded that our developed gave compatible result with IFA. Additionally, Our LAT are simple, rapid and suitable for detection in the field. However, for better sensitivity, diagnostic specificity, positive predictive value, negative predictive value, accuracy and Cohen's kappa comparison should be done in larger amounts of sera samples.
PubMed: 30733758
DOI: 10.1016/j.jgeb.2018.10.002 -
The Tohoku Journal of Experimental... Apr 2023The incidence of Brucella canis (B. canis) in humans is unknown in Northern Cyprus. In this study, we investigated the prevalence of B. canis and Brucella abortus (B....
The incidence of Brucella canis (B. canis) in humans is unknown in Northern Cyprus. In this study, we investigated the prevalence of B. canis and Brucella abortus (B. abortus) infection in human sera and evaluated the results obtained by agglutination-based techniques using standardized antigens made from B. canis comparatively. All of the subjects were negative in terms of Rose-Bengal plate test. Undiluted serum samples were initially screened by rapid slide agglutination test (RSAT), and those which were found positive were retested in the dilution of 1/25-1/200. Confirmation of the positive results was performed by using 2-mercaptoethanol standard agglutination test (SAT). The test antigen was prepared from the less mucoid M (-) variant of B. canis, and 1/1,048 titered dog antiserum was used as positive control. In 225 serum samples, 3.6% (8/225) was positive by B. canis M (-) RSAT, 4.4 % (10/225) was positive by B. canis M (-) indirect enzyme-linked immunosorbent assay (iELISA). 5.3% (12/225) was positive by B. abortus S99 RSAT and 9.8% (22/225) was positive by B. abortus S99 iELISA. Nine samples were positive by both B. abortus S99 RSAT and B. abortus S99 iELISA. Seven samples were positive by both B. canis M (-) RSAT and B. canis M (-) iELISA. One patient was positive by all methods. It is important to evaluate patient samples with RSAT and iELISA. Until the notification system gives better results to the Ministry of Health, in order to reach the real data for Northern Cyprus, multicenter prevalence determination studies should be done for future.
Topics: Humans; Animals; Dogs; Brucella canis; Brucella abortus; Brucellosis; Cyprus; Antigens, Bacterial; Antibodies, Bacterial; Enzyme-Linked Immunosorbent Assay; Agglutination Tests
PubMed: 36384858
DOI: 10.1620/tjem.2022.J096 -
Journal of Infection and Public Health Mar 2023Aim to investigate the brucella culture characteristics, diagnosis methods, and clinical characteristics, to provide the laboratory with diagnostic methods and...
OBJECTIVE
Aim to investigate the brucella culture characteristics, diagnosis methods, and clinical characteristics, to provide the laboratory with diagnostic methods and prevention and treatment for brucellosis.
METHODS
Data of 328 cases of brucellosis from 2012 to 2022 was analyzed, retrospectively. The bacterial culture characteristics, the clinical diagnostic methods, and the complications were analyzed respectively. The infection biomarkers of the brucellosis were analyzed by Receiver operating characteristic curve ROC.
RESULTS
Among the 328 brucellosis, 78.96 % of cases were men, the median age of the patients was (45.21±13.49) years and the annual incidence in our region was 67/100 000 per year. The diagnostic methods included pathogenic bacteria culture, serological diagnosis, and suspect case were 24.39 %, 47.56 %, and 28.05 %, respectively, sensitivity of combined detection Standard agglutination test (SAT) and the Rose Bengal test (RBT) is 96.2 %. In our work, 80 cases of brucellosis were diagnosed by a bacterial culture which were been identified as Brucella melitensis, blood culture was the main method (78.75 %) and the average positive alarm time was 80.74 (21.6-129) h and all of them were detected in aerobic bottles, followed by synovial fluid, bone marrow, lumbar spine, and joint tissue, puncture fluid and ascites culture which were 6.25 %, 3.75 %, 5.00 %, 5.00 % and 1.25 % respectively. The brucellosis with complications was lumbar spine lesions at 41.46 % cervical spine lesions at 4.60 % and knee joint lesions at 12.8 % and another osteoarthritis. The in-hospital mortality rate of the patients was 0.91 % and all of them were meningitis patients. ROC analysis indicated CRP had high sensitivity and specificity for brucellosis, and when CRP was 1.23mg/ml, the sensitivity and specificity were 0.727 and 0.718 respectively, and the U test also indicated CRP had a significant difference, Z=5.054, p <0.001.
CONCLUSIONS
Brucellosis is frequently morbidity in 40 + age men, which has been diagnosed by aerobic blood culture, generally bacterial culture, RBT and SAT, epidemiological, and commonly with complications of spine and arthropathy.
Topics: Adult; Female; Humans; Male; Middle Aged; Agglutination Tests; Antibodies, Bacterial; Brucella melitensis; Brucellosis; Retrospective Studies; Rose Bengal; Sensitivity and Specificity
PubMed: 36641837
DOI: 10.1016/j.jiph.2023.01.002 -
BMC Infectious Diseases Mar 2021Recent seroepidemiological studies have suggested that tularemia could be an endemic bacterial zoonosis in Iran.
BACKGROUND
Recent seroepidemiological studies have suggested that tularemia could be an endemic bacterial zoonosis in Iran.
METHODS
From January 2016 to June 2018, disease cases characterized by fever, cervical lymphadenopathy and ocular involvement were reported in Youzband Village of Kaleybar County, in the East Azerbaijan Province, northwestern Iran. Diagnostic tests included Francisella tularensis serology (including tube agglutination test and ELISA), PCR, and culture.
RESULTS
Among 11 examined case-patients, the tularemia tube agglutination test was positive in ten and borderline in one. PCR detected the F. tularensis ISFtu2 elements and fopA gene in one rodent and a spring water sample from the same geographic area.
CONCLUSIONS
Based on the clinical manifestations of the disease suggesting an oropharyngeal form of tularemia, serology results in case patients, and F. tularensis detection in the local fauna and aquatic environment, the water supply of the village was the likely source of the tularemia outbreak. Intervention such as dredging and chlorination of the main water storage tank of the village and training of villagers and health care workers in preventive measures and treatment of the illness helped control the infection.
Topics: Adolescent; Adult; Aged; Agglutination Tests; Animals; Bacterial Outer Membrane Proteins; Child; Child, Preschool; DNA, Bacterial; Female; Francisella tularensis; Fresh Water; Humans; Iran; Male; Mice; Polymerase Chain Reaction; Tularemia
PubMed: 33789598
DOI: 10.1186/s12879-021-06004-y -
Emerging Microbes & Infections Dec 2023Balancing the potentially serious outcomes of asymptomatic brucellosis and "waiting" for treatment in clinical practice is an urgent issue. Therefore, we assessed the... (Meta-Analysis)
Meta-Analysis
Balancing the potentially serious outcomes of asymptomatic brucellosis and "waiting" for treatment in clinical practice is an urgent issue. Therefore, we assessed the follow-up outcomes and epidemiological characteristics of asymptomatic brucellosis in the absence of treatment to provide evidence-based clinical clues. We searched eight databases in which 3610 studies from 1990 to 2021 were related to the follow-up outcomes of asymptomatic brucellosis. Thirteen studies, involving 107 cases, were finally included. Regarding the follow-up outcomes, we examined the presence or absence of symptoms and decreased serum agglutination test (SAT) titre. During the 0.5-18 months follow-up period, the pooled prevalence of appearing symptomatic was 15.4% (95% CI 2.1%-34.3%), cases that remained asymptomatic were 40.3% (95% CI 16.6%-65.8%), and decreased SAT titre was observed in 36.5% (95% CI 11.6%-66.1%). Subgroup analysis indicated that the pooled prevalence of appearing symptomatic with follow-up times of less than 6 months, 6-12 months, and 12-18 months was 11.5%, 26.4%, and 47.6%, respectively. The student subgroup had a higher prevalence of symptoms (46.6%) than the occupational and family populations. In conclusion, asymptomatic brucellosis has a high likelihood of appearing symptomatic and its severity may be underestimated. Active screening of occupational and family populations should be enhanced, and special attention should be paid to high-titre students for early intervention, if necessary. Additionally, future prospective, long-term, and large-sample follow-up studies are essential.
Topics: Humans; Follow-Up Studies; Brucellosis; Agglutination Tests; Prevalence
PubMed: 36849445
DOI: 10.1080/22221751.2023.2185464 -
Infectious Diseases of Poverty Mar 2021Brucellosis is an infectious-allergic zoonotic disease caused by bacteria of the genus Brucella. Early diagnosis is the key to preventing, treating, and controlling...
BACKGROUND
Brucellosis is an infectious-allergic zoonotic disease caused by bacteria of the genus Brucella. Early diagnosis is the key to preventing, treating, and controlling brucellosis. Fluorescence polarization immunoassay (FPA) is a new immunoassay for relatively rapid and accurate detection of antibodies or antigens based on antigen-antibody interaction. However, there is no report on FPA-based detection of human brucellosis in China. Therefore, this study is to evaluate the value of FPA for the diagnosis of human brucellosis in China.
METHODS
We recruited 320 suspected brucellosis cases who had the clinical symptoms and epidemiological risk factors between January and December, 2019. According to China Guideline for Human Brucellosis Diagnosis, the Rose Bengal test (RBT) was used for the screening test, and the serum agglutination test (SAT) was used as the confirmatory test. Brucellosis was confirmed only if the results of both tests were positive. Additionally, FPA and enzyme linked immune sorbent assay (ELISA) were compared with SAT, and their sensitivity, specificity, coincidence rate and consistency coefficient (Kappa value) as diagnostic tests were analyzed individually and in combination. The optimal cut-off value of FPA was also determined using the receiver operator characteristic (ROC) curve.
RESULTS
The optimum cut-off value of FPA was determined to be 88.5 millipolarization (mP) units, with a sensitivity of 94.5% and specificity of 100.0%. Additionally, the coincidence rate with the SAT test was 96.6%, and the Kappa value (0.9) showed excellent consistency. The sensitivity and specificity of FPA and ELISA combined were higher at 98.0% and 100.0% respectively.
CONCLUSIONS
When the cut-off value of FPA test is set at 88.5 mP, it has high value for the diagnosis of brucellosis. Additionally, when FPA and ELISA are combined, the sensitivity of diagnosis is significantly improved. Thus, FPA may have potential in the future as a diagnostic method for human brucellosis in China.
Topics: Agglutination Tests; Antibodies, Bacterial; Brucella; Brucellosis; Enzyme-Linked Immunosorbent Assay; Fluorescence Polarization Immunoassay; Humans; ROC Curve; Sensitivity and Specificity
PubMed: 33789762
DOI: 10.1186/s40249-021-00834-3 -
JGH Open : An Open Access Journal of... Jun 2021antibody levels in the blood are currently measured using an ELISA. In April 2016, FUJIFILM Wako Pure Chemical Corporation launched the "l-type Wako antibody J" test,...
BACKGROUND AND AIM
antibody levels in the blood are currently measured using an ELISA. In April 2016, FUJIFILM Wako Pure Chemical Corporation launched the "l-type Wako antibody J" test, which is based on the latex agglutination turbidimetric immunoassay. In this study, we investigated the usefulness of the Wako test.
METHODS
We measured antibody levels using both the ELISA and Wako test in 180 patients who underwent upper gastrointestinal endoscopy at our hospital between September 2017 and February 2019. Ninety patients were infected with . We calculated the diagnostic accuracy, sensitivity, and specificity of each test and the concordance rate between the ELISA and Wako test. If lower limits of 90% confidence intervals (CIs) for each diagnostic validity exceeded the 85% threshold, the usefulness of the diagnostic test was confirmed.
RESULTS
The diagnostic accuracy, sensitivity, and specificity were 94.4% (90% CI, 90.8-97.0%), 94.4% (90% CI, 88.7-97.8%), and 94.4% (90% CI, 88.7-97.8%), respectively, when the Wako test was used, and 94.4% (90% CI, 90.8-97.0%), 88.9% (90% CI, 81.9-93.8%), and 100% (90% CI, 96.0-100%), respectively, when the ELISA was used. The concordance rate between the two tests was high ( = 0.8444).
CONCLUSIONS
We confirmed the usefulness of the Wako test, especially when screening for infection, due to its high sensitivity.
PubMed: 34124385
DOI: 10.1002/jgh3.12553 -
Journal of Clinical Laboratory Analysis May 2020Rotavirus A and human adenovirus are the two most common causes of infantile diarrhea; thus, it is of great importance to find out a rapid and accurate diagnostic...
OBJECTIVES
Rotavirus A and human adenovirus are the two most common causes of infantile diarrhea; thus, it is of great importance to find out a rapid and accurate diagnostic method. This study aimed to evaluate the diagnostic significance of latex agglutination test for detection of rotavirus A and human adenovirus.
METHODS
A prospective study was conducted on 214 diarrhea children from September 2018 to March 2019 in our hospital. Fresh stool samples were collected for detection of rotavirus A and human adenovirus by latex agglutination test and quantitative reverse transcription polymerase chain reaction (RT-qPCR). Then, the consistency of results detected by these two methods was analyzed. RESULTS: With performing the latex agglutination test, it was revealed that positive rates for detecting rotavirus A virus and human adenovirus were 23.83% (51/214) and 25.24% (54/214), respectively. Meanwhile, results of RT-qPCR showed that positive rates for detecting rotavirus A virus and human adenovirus were 58 (27.10%) and 59 (27.57%), respectively. Using RT-qPCR as the gold standard, the sensitivity and specificity of the latex agglutination test for detecting rotavirus A were 81.03% and 97.44%, and the corresponding values for detecting human adenovirus were 76.27% and 94.19%, respectively.
CONCLUSION
This latex agglutination test showed a satisfactory consistency with RT-qPCR for detecting rotavirus A and human adenovirus. The mentioned commercial assay may be highly appropriate for rapid screening of rotavirus A and human adenovirus.
Topics: Adenovirus Infections, Human; Adenoviruses, Human; Child, Preschool; Diarrhea; Feces; Humans; Latex Fixation Tests; Prospective Studies; Reverse Transcriptase Polymerase Chain Reaction; Rotavirus; Rotavirus Infections
PubMed: 31930752
DOI: 10.1002/jcla.23208 -
Sexually Transmitted Diseases Jul 2019Syphilis transmission can be prevented by prompt diagnosis and treatment of primary and secondary infection. We evaluated the performance of a point-of-care rapid...
BACKGROUND
Syphilis transmission can be prevented by prompt diagnosis and treatment of primary and secondary infection. We evaluated the performance of a point-of-care rapid syphilis treponemal (RST) test in an emergency department (ED) setting.
METHODS
Between June 2015 and April 2016, men aged 18 to 34 years seeking services in a Detroit ED, and with no history of syphilis, were screened for syphilis with the RST test, rapid plasma reagin (RPR) test, and Treponema pallidum particle agglutination assay (TP-PA). A positive reference standard was both a reactive RPR and a reactive TP-PA. We compared test results in self-reported men who have sex with men (MSM) to non-MSM.
RESULTS
Among 965 participants, 10.9% of RST tests were reactive in MSM and only 1.5% in non-MSM (P < 0.001). Sensitivity of the RST test was 76.9% and specificity was 99.0% (positive predictive value, 50.0%) compared with the positive reference standard. Three discordant specimens found negative with the RST test but positive with the reference standard had an RPR titer of 1:1, compared with 10 specimens with concordant positive results that had a median RPR titer of 1:16. The RST sensitivity was 50.0% (positive predictive value, 68.4%) compared to the TP-PA test alone. Among men seeking care in an ED, the RST detected 76.9% of participants with a reactive RPR and TP-PA.
CONCLUSIONS
The RST test detected all of the participants with an RPR titer ≥1:2 but less than 20% of participants with a positive TP-PA and negative RPR. The RST test was useful to detect a high proportion of participants with an active syphilis in an urban ED.
Topics: Adolescent; Adult; Agglutination Tests; Antibodies, Bacterial; Emergency Service, Hospital; Humans; Male; Michigan; Point-of-Care Testing; Reagins; Sensitivity and Specificity; Sexual and Gender Minorities; Syphilis; Syphilis Serodiagnosis; Time Factors; Treponema pallidum; Young Adult
PubMed: 30839394
DOI: 10.1097/OLQ.0000000000000993 -
BMC Research Notes Jul 2021Canine visceral leishmaniasis (CVL) is the main source of human visceral leishmaniosis (HVL) in Mediterranean region, including Iran and is spread from domestic dogs to...
OBJECTIVE
Canine visceral leishmaniasis (CVL) is the main source of human visceral leishmaniosis (HVL) in Mediterranean region, including Iran and is spread from domestic dogs to Phlebotomine sand flies vectors to humans. To control the transmission of HVL, early and accurate detection of infected dogs is paramount importance despite it remains a confronting challenge. Herein, we evaluated the performance of direct agglutination test (DAT) against gold standard nested polymerase chain reaction (nested-PCR) for CVL diagnosis in symptomatic and asymptomatic domestic dogs from endemic areas of Iran.
RESULTS
Venous blood samples were collected from dogs without clinical signs (n = 30) and with clinical signs (n = 35) suggestive of Leishmania infantum infection. Among 65 samples examined, Leishmania DNA was detected by nested-PCR in 89.23% (58/65). Furthermore, 86.15% (56/65) nested-PCR positive samples were also DAT positive. The results of the DAT sensitivity test were 96.43% and 96.67% in symptomatic and asymptomatic dogs, respectively, while the specificity was 100.00% and 60.00% in symptomatic and asymptomatic dogs, respectively. The results of this study also pointed out substantial concordance between DAT test and nested-PCR method in both symptomatic dogs (Κ = 0.783; P < 0.001) and asymptomatic dogs (Κ = 0.618; P < 0.001). Thus, DAT represents as a simple and economic tool for initial diagnosis of CVL particularly in endemic areas of the disease.
Topics: Agglutination Tests; Animals; Antibodies, Protozoan; DNA, Protozoan; Dog Diseases; Dogs; Iran; Leishmania infantum; Leishmaniasis, Visceral; Polymerase Chain Reaction
PubMed: 34256817
DOI: 10.1186/s13104-021-05654-0