scholarly journals Challenges and new methods in the diagnosis of Lyme disease in children

2020 ◽  
pp. 652-661
Keyword(s):  
Lyme Disease ◽  
Clinical Symptoms ◽  
Direct Detection ◽  
Serological Test ◽  
Direct Methods ◽  
Disease Diagnosis ◽  
Detection Methods ◽  
Laboratory Diagnostics ◽  
Diagnostic Strategies ◽  
Culture Techniques

Despite the constant development of biotechnology, laboratory diagnostics of Lyme disease in children still poses a significant challenge. The aim of this article is to present the current methods of Lyme disease diagnosis and its future perspectives. A serological test is often the first step in supporting clinical diagnosis of Lyme disease in children. Recently, a new generation of enzyme-linked immunosorbent assays has been created. These assays use recombinant proteins or synthetic peptides in their antigenic spectrum. It is postulated that these tests may replace the classic immunoblot as the second step in the Lyme disease diagnostic protocol. Direct detection methods based on bacterial culture techniques or using the polymerase chain reaction (PCR) have inadequate sensitivity, which prevents their widespread use in clinical practice. Recently, a number of other tools have been developed that are of supportive importance. Among them, measuring of the CXCL13 chemokine concentration in the cerebrospinal fluid has the potential to become a routine procedure in the diagnosis of Lyme disease in children. Future diagnostic strategies of Lyme disease might include: innovative immunological tests using new antigens, combining serology with direct methods in order to increase sensitivity, standardization of selected unconventional tests, identification of host response biochemical metabolic markers or linking clinical symptoms reported by patients with appropriate test panels. In the absence a vaccine which protects against the disease, the preventive recommendations given to parents to prevent tick bites in children remain valid.

scholarly journals Expanding Access to Biospecimens for Lyme Disease Test Development

2020 ◽  
Vol 58 (6) ◽  
Author(s):  
John L. Schmitz
Keyword(s):  
Lyme Disease ◽  
Whole Blood ◽  
Direct Detection ◽  
Test Development ◽  
Diagnostic Testing ◽  
Nucleic Acid Amplification ◽  
Detection Methods ◽  
Antibody Detection ◽  
Laboratory Diagnostics ◽  
Content Type

ABSTRACT The laboratory diagnosis of Lyme disease relies upon serologic testing. A standard or modified two-tiered testing algorithm is used to enhance the accuracy of antibody detection. However, this approach suffers from a lack of sensitivity in early Lyme disease. Ongoing efforts to develop more sensitive antibody detection technologies and other diagnostic approaches are dependent upon the availability of quality-assured biospecimens linked to reliable clinical data. In this issue of the Journal of Clinical Microbiology, Horn et al. (E. J. Horn, G. Dempsey, A. M. Schotthoefer, U. L. Prisco, et al., J Clin Microbiol 58:e00032-20, 2020, https://doi.org/10.1128/JCM.00032-20) described the development of the Lyme Disease Biobank. Clinically categorized case patients with early Lyme disease and healthy controls were identified (without laboratory diagnostic testing) from three sites where Lyme disease is endemic. Subjects provided whole blood and urine, which were processed and stored at a central biorepository. Whole blood, serum, and urine aliquots were prepared and are available to investigators developing laboratory diagnostics for Lyme disease. After obtaining samples, extensive laboratory testing was performed, including serologic and nucleic acid amplification testing for B. burgdorferi and other tick-borne pathogens. Direct detection methods yielded few positive results. Relative to the findings for another commonly used biorepository cohort, the results of this testing demonstrated a low seropositive rate, as determined by standard two-tiered testing. Additionally, relatively few subjects demonstrated seroconversion with testing of convalescent-phase samples. This clinical and serologically defined cohort of samples from Lyme disease and control cases from areas of Lyme disease endemicity offers an additional valuable resource for novel test development that includes alternate specimen types.

scholarly journals Practical aspects of Lyme disease in children

2020 ◽  
pp. 33-38
Author(s):  
N.V. Banadyha ◽  
◽  
I.O. Rogalskyy ◽  
Keyword(s):  
Lyme Disease ◽  
Lyme Borreliosis ◽  
Conflict Of Interest ◽  
Patient Management ◽  
Clinical Symptoms ◽  
Clinical Manifestations ◽  
Laboratory Diagnostics ◽  
Pediatric Practice ◽  
Key Words Lyme Disease ◽  
The Moment

Lyme disease is especially important in the spring–autumn period, despite the fact that its clinical manifestations may be throughout the year. Awareness of general practitioners with this problem is insufficient, it requires additional knowledge about diagnosis and treatment. In pediatric practice, the fact of a child being bitten by an Ixodes mite that carries the causative agent of Lyme borreliosis, in addition to babesiosis, anaplasmosis, often goes unnoticed. Therefore, parents seek medical help only when various problems arise, often the thought of Lyme disease does not even arise. Lyme disease has a wide polymorphism of clinical symptoms, is characterized by multisystem lesions, cyclical course — all this complicates the diagnostic search. The difficulty also lies in the fact that there are no domestic clinical recommendations. This publication presents approaches to the diagnosis, treatment, prevention of Lyme disease based on the experience of experts from different countries. Modern approaches to two-stage laboratory diagnostics, tactics of patient management from the moment of bite, treatment at different stages of Lyme borreliosis are analyzed. Attention is paid to the need for epidemiological research in Ukraine and the beginning of educational programs to prevent the disease. The differentiated approach to the treatment of Lyme disease in children due to age aspects, concomitant pathology, safety of long_term antibacterial therapy should be studied more. No conflict of interest was declared by the authors. Key words: Lyme disease, children, diagnosis, treatment, prevention.

scholarly journals Evaluation the efficiency of Trichomonas vaginalis depending on clinical sings , direct examination ,culturing and serological test

2011 ◽  
Vol 8 (1) ◽  
pp. 392-399
Author(s):  
Baghdad Science Journal
Keyword(s):  
Trichomonas Vaginalis ◽  
Clinical Symptoms ◽  
Culture Media ◽  
Serological Test ◽  
Clinical Signs ◽  
Direct Methods ◽  
Laboratory Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Indirect Haemagglutination ◽  
Number Of Patients

The adequacy of diagnostic tests, together with trichomoniasis associated clinical symptoms, were investigated in females suffering vaginitis, and they were referred to the Gynecology Department, Al-Yarmouk Teaching Hospital during the period December 2004 – June 2005. The total number of patients was 250 cases (age range: 18 - 52 years), and each patient was examined using a sterile speculum to obtain vaginal swabs for examination. The diagnosis with T. vaginalis was done in many methods. The direct methods included wet and stained (Leishman's stain) examinations and cultivation in different culture media (Kupferberg Trichomonas Broth Base;, Trichomonas Agar Base; TAB and Trichomonas Modified CPLM), while the indirect methods were serological detections of anti-trichomonas antibodies in the sera of patients by using of indirect haemagglutination test (IHAT) and enzyme linked immunosorbent assay (ELISA). The results of this profile were as the following: 1. Out of 250 females with abnormal vaginal discharges, 15 patients (6%) were infected with T. vaginalis. 2. The sensitivity of testing methods was different. It was 66.7 and 13.3% for wet and stained examinations, respectively. In culture examinations, the sensitivity was 60, 80 and 100% for Kupferberg, TAB and CPLM media, respectively, while the sensitivity IHAT and ELISA were 40 and 73.3%, respectively. 3. The clinical sings of infection in women were discharge and itching (46.7%), discharge with itching and dysuria (20.0%), discharge only (13.3%), itching only (13.3%) and discharge with dysuria (6.7%). Therefore we can conclude that impossible to the clinical signs in diagnose of Trichomonasis because of the variation and the laboratory diagnosis is necessary especially the cultivation method.

scholarly journals A Case of Undetected Neuroborreliosis in a 75-Year-Old Chinese Male

2018 ◽  
Vol 2018 ◽  
pp. 1-4
Author(s):  
Jenny Lamichhane ◽  
Rohail Haider ◽  
Michael Bekkerman ◽  
Sean Tilford ◽  
Shireen Hijaz ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Clinical Symptoms ◽  
Sensu Stricto ◽  
Blood Cultures ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cerebrospinal Fluid Analysis ◽  
Culture Techniques ◽  
Fluid Analysis ◽  
Chinese Male ◽  
Unstable Gait

Lyme disease is a multisystem infection caused by the spirochete Borrelia burgdorferi sensu stricto that manifests with characteristic symptoms in patients. Patients are identified based on their clinical symptoms and then diagnosed through enzyme-linked immunosorbent assay (ELISA), Western blot, and blood culture techniques. Here, we present the case of a 75-year-old, Northeast suburban resident complaining of unstable gait, high fevers, malaise, myalgia, and confusion. This patient’s symptoms were nonspecific, and his lab titers and blood cultures were repeatedly negative during his stay. It was only late in the course of his treatment that blood titers and cerebrospinal fluid analysis were positive for Lyme IgG and IgM. He was treated with intravenous doxycycline and prescribed oral doxycycline on discharge, resulting in a full recovery. We express the need for physicians to consider Lyme disease in endemic patients presenting with nonspecific systemic signs.

scholarly journals Comparison of SARS-CoV-2 Indirect and Direct Detection Methods

2020 ◽  
Author(s):  
Joel D. Pearson ◽  
Daniel Trcka ◽  
Sharon J. Hyduk ◽  
Marie-Ming Aynaud ◽  
J. Javier Hernández ◽  
...  
Keyword(s):  
Direct Detection ◽  
Detection System ◽  
Rapid Development ◽  
Rna Extraction ◽  
Direct Methods ◽  
Rna Isolation ◽  
Clinical Results ◽  
Detection Methods ◽  
Indirect Methods ◽  
Detection Systems

ABSTRACTThe COVID-19 pandemic caused by the SARS-CoV-2 virus has placed extensive strain on RNA isolation and RT-qPCR reagents. Rapid development of new test kits has helped to alleviate these shortages. However, comparisons of these new detection systems are largely lacking. Here, we compare indirect methods that require RNA extraction, and direct RT-qPCR on patient samples. For RNA isolation we compared four different companies (Qiagen, Invitrogen, BGI and Norgen Biotek). For detection we compared two recently developed Taqman-based modules (BGI and Norgen Biotek), a SYBR green-based approach (NEB Luna Universal One-Step Kit) with published and newly-developed primers, and clinical results (Seegene STARMag RNA extraction system and Allplex 2019-nCoV RT-qPCR assay). Most RNA isolation procedures performed similarly, and while all RT-qPCR modules effectively detected purified viral RNA, the BGI system proved most sensitive, generating comparable results to clinical diagnostic data, and identifying samples ranging from 65 copies – 2.1×105 copies of viral Orf1ab/μl. However, the BGI detection system is ∼4x more expensive than other options tested here. With direct RT-qPCR we found that simply adding RNase inhibitor greatly improved sensitivity, without need for any other treatments (e.g. lysis buffers or boiling). The best direct methods were ∼10 fold less sensitive than indirect methods, but reduce sample handling, as well as assay time and cost. These studies will help guide the selection of COVID-19 detection systems and provide a framework for the comparison of additional systems.

scholarly journals A Novel Design Combining Isothermal Exponential Amplification and Gold-Nanoparticles Visualization for Rapid Detection of miRNAs

2018 ◽  
Vol 19 (11) ◽  
pp. 3374 ◽  
Author(s):  
Jiquan Jiang ◽  
Bin Zhang ◽  
Chi Zhang ◽  
Yifu Guan
Keyword(s):  
Early Phase ◽  
Color Change ◽  
Direct Detection ◽  
Colorimetric Assay ◽  
Disease Diagnosis ◽  
Dna Probes ◽  
Time Saving ◽  
Promising Alternative ◽  
Sandwich Hybridization ◽  
Wide Range

MicroRNAs (miRNAs) play important roles in a wide range of biological processes, and their aberrant expressions are associated with various diseases. The levels of miRNAs can be useful biomarkers for cellular events or disease diagnosis; thus, sensitive and selective detection of microRNAs is of great significance in understanding biological functions of miRNAs, early-phase diagnosis of cancers, and discovery of new targets for drugs. However, traditional approaches for the detection of miRNAs are usually laborious and time-consuming, with a low sensitivity. Here, we develop a simple, rapid, ultrasensitive colorimetric assay based on the combination of isothermal Exponential Amplification Reaction (EXPAR) and AuNP-labeled DNA probes for the detection of miRNAs (taking let-7a as a model analyte). In this assay, the presence of let-7a is converted to the reporter Y through EXPAR under isothermal conditions. The subsequent sandwich hybridization of the reporter Y with the AuNP-labeled DNA probes generates a red-to-purple color change. In other words, if the reporter Y is complementary to the AuNP-labeled DNA probes, the DNA-functionalized AuNPs will be aggregated, resulting in the change of solution color from red to purple/blue, while when the AuNP-labeled DNA probes are mismatched to the reporter Y, the solution remains red. This assay represents a simple, time-saving technique, and its results can be visually detected with the naked eye due to the colorimetric change. The method provides superior sensitivity, with a detection limit of 4.176 aM over a wide range from 1 nM to 1 aM under optimal conditions. The method also shows high selectivity for discriminating even single-nucleotide differences between let-7 miRNA family members. Notably, it is comparable to the most sensitive method reported to date, thus providing a promising alternative to standard approaches for the direct detection of let-7a miRNA. Importantly, through combination with specific templates, different miRNAs can be converted to the same reporter Y, which can hybridize with the same set of AuNP-labeled DNA probes to form sandwich hybrids. The color change of the solution can be observed in the presence of the target miRNA. This technique has potential as a routine method for assessing the levels of miRNAs, not only for let-7, but also for various miRNAs in the early phase of cancers. In addition, it can be a useful tool in biomedical research and clinical diagnosis, as well as diagnosis or surveillance programs in field conditions.

scholarly journals The Relapsing Fever Spirochete Borrelia hermsiiContains Multiple, Antigen-Encoding Circular Plasmids That Are Homologous to the cp32 Plasmids of Lyme Disease Spirochetes

2000 ◽  
Vol 68 (7) ◽  
pp. 3900-3908 ◽  
Author(s):  
Brian Stevenson ◽  
Stephen F. Porcella ◽  
Katrina L. Oie ◽  
Cecily A. Fitzpatrick ◽  
Sandra J. Raffel ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Direct Detection ◽  
Human Infection ◽  
Relapsing Fever ◽  
Borrelia Hermsii ◽  
Functional Studies ◽  
Dna Library ◽  
Multiple Antigen ◽  
Antigenic Proteins

ABSTRACT Borrelia hermsii, an agent of tick-borne relapsing fever, was found to contain multiple circular plasmids approximately 30 kb in size. Sequencing of a DNA library constructed from circular plasmid fragments enabled assembly of a composite DNA sequence that is homologous to the cp32 plasmid family of the Lyme disease spirochete,B. burgdorferi. Analysis of another relapsing fever bacterium, B. parkeri, indicated that it contains linear homologs of the B. hermsii and B. burgdorfericp32 plasmids. The B. hermsii cp32 plasmids encode homologs of the B. burgdorferi Mlp and Bdr antigenic proteins and BlyA/BlyB putative hemolysins, but homologs of B. burgdorferi erp genes were absent. Immunoblot analyses demonstrated that relapsing fever patients produced antibodies to Mlp proteins, indicating that those proteins are synthesized by the spirochetes during human infection. Conservation of cp32-encoded genes in differentBorrelia species suggests that their protein products serve functions essential to both relapsing fever and Lyme disease spirochetes. Relapsing fever borreliae replicate to high levels in the blood of infected animals, permitting direct detection and possible functional studies of Mlp, Bdr, BlyA/BlyB, and other cp32-encoded proteins in vivo.

scholarly journals Laboratory Diagnostics Performance in Uganda: A Survey of Test Availability and Constraints Across 100 Laboratories

2021 ◽  
Author(s):  
Noel Namuhani ◽  
Suzanne N Kiwanuka ◽  
Martha Akulume ◽  
Simeon Kalyesubula ◽  
William Bazeyo ◽  
...  
Keyword(s):  
Clinical Laboratory ◽  
Disease Diagnosis ◽  
Sub Saharan Africa ◽  
Essential Medicines ◽  
Full Blood Count ◽  
Laboratory Diagnostics ◽  
Cross Sectional ◽  
Laboratory Services ◽  
Control And Prevention ◽  
Sub Saharan

Abstract Background Clinical laboratory services are a critical component of the health system for effective disease diagnosis, treatment, control and prevention. However, many laboratories in Sub Saharan Africa remain dysfunctional. The high costs of tests in the private sector also remain a hindrance to accessing testing services. This study aimed at assessing the functionality of laboratories based on test menus and the associated constraints in Uganda. Methods This cross sectional quantitative study involved an assessment of 100 laboratories randomly selected in 20 districts from four regions of the country. Sixteen percent of the studied laboratories were regional hub laboratories. Laboratory in charges and managers in each of the selected laboratories were interviewed. A checklist for laboratory supplies adapted from the Essential Medicines and Health supplies list for Uganda, (2012) was used to assess availability of testing supplies. Data was analyzed using excel and STATA 14. Results At the point of assessment, generally, all laboratories were able to perform malaria tests and HIV tests. All the hub laboratories conducted malaria tests and TB screening. Less than half had electrolytes tests due to lack of equipment, nonfunctioning equipment and lack of reagents. Full blood count tests were missing in 25% of the hub laboratories mainly due to lack of equipment. The lack of reagents (66.7%) and the lack of equipment (58.3%) caused the majority 10/16 of the hubs to routinely referred specimens for tests that are supposed to be carried out in these laboratories due to lack of reagents (66.7%) and non-functional equipment (58.3%). Although officially recognized as an operational structure, Hub laboratories lacked a list of essential and vital supplies. Conclusions Most laboratories performed well for the common tests. However, many laboratories did not meet testing requirements especially for the advanced tests according to standard testing menus for Uganda due to non-functioning equipment, lack of equipment and reagents. Hubs lack list of essential supplies. Therefore, there is need to provide equipment to laboratories, repair the non-functional ones and develop an essential list of supplies for the hub laboratories.

scholarly journals Some Predictors of Depressive Disorders in the Context of Clinical Patomorphosis

2021 ◽  
Vol 6 (4) ◽  
pp. 77-82
Author(s):  
O. O. Belov ◽  
Keyword(s):  
Cognitive Impairment ◽  
Clinical Picture ◽  
Depressive Disorders ◽  
Clinical Symptoms ◽  
Emotional Lability ◽  
Obsessive Compulsive ◽  
Diagnostic Strategies ◽  
Low Mood ◽  
Self Blame ◽  
Initial Stage

The purpose of the study was to study the clinical and psychopathological phenomenology of the initial stage of depressive disorders in the context of clinical pathomorphosis. Materials and methods. Features of clinical symptoms of the initial stage of depressive disorders in the comparative aspect in the context of clinical pathomorphosis based on the analysis of medical records of 236 patients who were treated for depressive disorders in 1971-1995 (ICD-9 codes 296.1, 296.3) and clinical examination of 245 patients with depressive disorders in 2015-2019 (ICD-10 codes F 31.3, F 31.4, F 32.0, F 32.1, F 32.2, F 33.0, F 33.1, F 33.2) are considered. Results and discussion. It was established that there is a predominance in the clinical picture of modern depressive disorders of low mood (in general in 91.4% of patients, 91.6% of men and 91.3% of women, p>0.05), dyssomnia (93.1%, 92.5% and 93.5%, respectively, p>0.05), anxiety, fear (84.5%, 78.5%, 89.1%, respectively, p<0.01), asthenia (82.4%, 77.6% and 86.2%, respectively, p>0.05), somatic vegetative symptoms (82.9%, 77.6% and 87.0%, respectively, p<0.01), apathy (78.8%, 69.2% and 86.2%, respectively, p<0.01) and ideas of self-humiliation and self-blame (69.8%, 72.9% and 67.4%, respectively, p<0.01), and the relatively low prevalence of obsessive symptoms (55.1%, 54.2% and 55.8%, respectively, p<0.05), emotional lability (51.0%, 54.2% and 48.6%, respectively, p<0.01) and cognitive impairment (45.3%, 43.9% and 46.4%, respectively, p<0.05) with a predominance of emotional lability and ideas of self-humiliation and self-blame in men, and manifestations of anxiety, fear, apathy, cognitive impairment, obsessive and somatic vegetative symptoms in women, which gives grounds to consider that the main predictors of depressive disorder at the initial stage of low mood are dyssomnia, anxiety fear, asthenia and somatic vegetative symptoms. The revealed features suggest the presence of a clinical pathomorphosis of depressive disorders. The clinical pathomorphosis of the initial stage of depressive disorders is in a significant reduction in the clinical picture of low mood, ideas of self-abasement and self-blame, emotional lability and cognitive impairment, and an increase in anxiety, fear, asthenia, apathy, obsessive symptoms and obsessive-compulsive symptoms, with significantly greater gender differentiation of clinical symptoms of depression. Conclusion. The identified patterns are embedded in the general trend towards polymorphism and clinical undifferentiation of modern depressive disorders, significant involvement of patients with sleep disorders, asthenic, apathetic and somatic vegetative symptoms, which requires revision of diagnostic strategies and individualization of diagnosis. The identified patterns can be used for early diagnosis of depressive disorders and prevention of depression

scholarly journals Clinical and epidemiological characteristics of tick-borne borreliosis in the Krasnodar Krai

2014 ◽  
Vol 19 (1) ◽  
pp. 4-11
Author(s):  
M. G. Avdeeva ◽  
D. Yu. Moshkova ◽  
L. P. Blazhnyaya ◽  
V. N. Gorodin ◽  
S. V Zotov ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Clinical Symptoms ◽  
Early Period ◽  
Epidemiological Data ◽  
Course Of Disease ◽  
Krasnodar Krai ◽  
Natural Foci ◽  
The Republic ◽  
Epidemiological Characteristics ◽  
The City

The purpose of the study the improvement of early diagnostics of Lyme disease on the based of clinical and epidemiological analysis of the main clinical forms of the acute course of the disease in new natural foci in the Krasnodar Krai. Patients and methods. There was analyzed the clinical course of disease and epidemiological data for 207 patients in the early period of acute course of Lyme disease within the period from 2004 to 2013. Results. In the territory of the Krasnodar Krai and the Republic of Adygea there has formed a set of foci of tick-borne borreliosis (TB), i.e. Lyme disease (LD). The infection of cases is observed not only in the natural foci of disease, but also within the city limits. The disease is registered mainly in the erythematous form (74% of patients), non-erythematous form accounts for 26%. The average age of patients was 41,1±1,83, years, males 38%, females 62%. In the non-erythematous form initial clinical symptoms are recorded in average in 11,6 ± 2,20 days after tick suction; in the erythematous form in 6,4 ± 0,70 days ( p

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