Laboratory Diagnostic Procedures Research Articles

Exploring Acute Diarrhea in Infants: A Comprehensive Review of Infectious Agents and Laboratory Diagnostic Techniques

Introduction: acute diarrhea is one of the main causes of mortality in children under 5 years of age around the world. They disproportionately affect underdeveloped countries, where access to health services, drinking water and sanitation is limited. Objective: this review identifies the most relevant and recent studies associated with the topic, recognizes the most common infectious agents and determines the laboratory techniques used for their diagnosis. Methods: the literature was reviewed from sources such as Elsevier, PubMed, Scopus, SciELO and Google Scholar during the period March-May 2024, published in the last 5 years and several studies prior to 2019 that contributed relevant information to the topic. Results: previous studies reveal that acute childhood diarrhea is mainly characterized by its fecal-oral transmission, caused by Rotavirus and Adenovirus, and sometimes by Escherichia coli, Salmonella spp., Giardia lamblia and Entamoeba histolytica, among others. Acute diarrhea is characterized by the presence of three or more soft or liquid stools per day, and is classified according to its duration. Fecal increase, dehydration and electrolyte abnormalities are associated complications and constitute clinically useful elements for its diagnosis. For viruses, rapid tests and PCR are used, for bacteria, cultures and antibiograms, and for parasites, coproparasites, ELISA and PCR. Conclusion: These disorders represent a significant health challenge, with a considerable impact on morbidity and mortality, originating from viruses, bacteria and parasites. The diagnosis is made through physical, coprological examinations, immunochromatographic and molecular tests

Do dynamic changes in haematological and biochemical parameters predict mortality in critically ill COVID-19 patients?

BACKGROUND: Critically ill COVID-19 patients are usually subjected to clinical, laboratory, and radiological diagnostic procedures resulting in numerous findings. Utilizing these findings as indicators for disease progression or outcome prediction is particularly intriguing. OBJECTIVES: Exploring the significance of dynamic changes in haematological and biochemical parameters in predicting the mortality of critically ill COVID-19 patients. METHODS: The present study was a prospective and observational study involving mechanically ventilated 75 critically ill adult COVID-19 patients with hypoxemic respiratory failure. The collected data included baseline patient characteristics, treatment options, outcome, and laboratory findings at admission and 7 days after. The dynamics of the obtained findings were compared between survivors and non-survivors. RESULTS: The 28-day survival rate was 61.3%. In the group of non-survivors significant dynamic changes were found for C-reactive protein (p= 0.001), interleukin-6 (p< 0.001), lymphocyte (p= 0.003), neutrophil-lymphocyte ratio (p= 0.003), platelets (p< 0.001), haemoglobin (p< 0.001), iron (p= 0.012), and total iron-binding capacity (p< 0.001). Statistically significant changes over time were found for ferritin (p= 0.010), D-dimer (p< 0.001), hs-troponin T (p< 0.002), lactate dehydrogenase (p= 0.001), glucose (p= 0.023), unsaturated iron-binding capacity (p= 0.008), and vitamin D (p< 0.001). CONCLUSION: The dynamic changes in inflammatory, haematological and biochemical parameters can predict disease severity, and outcome.

Analysis of thicknesses of blood collection needle by scanning electron microscopy reveals wide heterogeneity.

The preanalytical phase in clinical laboratory diagnostics is currently receiving more and more attention. This term describes one part of actions and aspects of the "brain-to-brain cycle" of the medical laboratory diagnostic procedure that take place before the analytical phase. However, the preanalytical activities, the handling of unsuitable samples and the reporting procedures are neither fully standardized nor harmonized worldwide. The influence of the properties of the blood collection needle must be acknowledged. In this work, we focused on the investigation of the internal structure and size of standardized 21G blood collection needles. All parameters were measured with a scanning electron microscope using a Jeol model JSM-6000PLUS. Our. The obtained data shows that the internal surfaces of the needles vary greatly from manufacturer to manufacturer (by around 35 %), and this may play an important role in influencing blood flow and even the risk of blood cell injury (especially hemolysis) during blood drawing. The differential actual needle diameters can vary greatly between needle manufactures and this variety may have a significant impact on laboratory values and may also lead to specimen rejection.

Clinical laboratory diagnostic techniques used in the identification of the helminth Ascaris lumbricoides, a review of the literature

Ascariasis is a disease caused by the helminth Ascaris lumbricoides (intestinal worm), which infects an average of 819 million individuals. Approximately more than 60 000 people die worldwide each year from this parasite. It is classified as a neglected subtropical endemic disease and is frequently found in Africa, Latin America, and East Asia. Ascariasis is detected by conventional tests, which are based on the direct identification of A. lumbricoides by direct macroscopic examination, traditional microscopy, formaldehyde concentration method, Kato-Katz, McMaster, flotation techniques (such as FLOTAC and Mini-FLOTAC), with the aim of observing helminth eggs; all these are cost-effective tests, but of low or moderate sensitivity. Therefore, molecular biology methods such as conventional PCR and multiplex PCR are an option for the detection of this parasite due to their high sensitivity and specificity, but their low accessibility in endemic countries due to the high prices of reagents and equipment should be considered. Indirect methods based on serology have been used for the identification of A. lumbricoides, but they have generated cross reactions with infections by other helminths so there is no specific serological test for this parasite at present. Nowadays, new techniques have been designed with high sensitivity and specificity that allow the diagnosis of ascariasis, among which we can mention detection and identification of A. lumbricoides eggs through optical imaging and loop-mediated isothermal amplification

Mastitis in goats – a still current problem

Even though goats are one of the first animal species domesticated by humans, udder inflammation in these female farm animals still poses major diagnostic and therapeutic challenges for practicing veterinarians. Mastitis in goats, similarly to cattle, causes large economic losses, including breeding losses and losses in the dairy industry. For this reason, the most important information regarding the diagnosis and treatment of mastitis in goats has been collected in this study, based on the latest data from the literature. Attention was drawn to the fact that in many forms of mastitis, to achieve the desired therapeutic effect it is crucial to undertake quick and targeted therapy. The study also emphasized the need to develop new laboratory diagnostic techniques aimed at determining indicators of the inflammatory reaction not only in milk and blood serum but also in saliva collected from females suffering from mastitis. This enables a quick and effective diagnosis of the disease and an effective treatment method.

Stability of African swine fever virus genome under different environmental conditions.

African swine fever (ASF), a globally transmitted viral disease caused by ASF virus (ASFV), can severely damage the global trade economy. Laboratory diagnostic methods, including pathogen and serological detection techniques, are currently used to monitor and control ASF. Because the large double-stranded DNA genome of the mature virus particle is wrapped in a membrane, the stability of ASFV and its genome is maintained in most natural environments. This study aimed to investigate the stability of ASFV under different environmental conditions from both genomic and antibody perspectives, and to provide a theoretical basis for the prevention and elimination of ASFV. In this study, we used quantitative real-time polymerase chain reaction for pathogen assays and enzyme-linked immunosorbent assay for serological assays to examine the stability of the ASFV genome and antibody, respectively, under different environmental conditions. The stability of the ASFV genome and antibody under high-temperature conditions depended on the treatment time. In the pH test, the ASFV genome and antibody remained stable in both acidic and alkaline environments. Disinfection tests revealed that the ASFV genome and antibody were susceptible to standard disinfection methods. Collectively, the results demonstrated that the ASFV genome is highly stable in favorable environments but are also susceptible to standard disinfection methods. This study focuses on the stability of the ASFV genome under different conditions and provides various standard disinfection methods for the prevention and control of ASF.

MICROBIAL PROFILE OF ULCERATIVE KERATITIS AT TERTIARY LEVEL HOSPITAL

Background: Keratitis is the term applied for inammations of the cornea. Ulcerative keratitis is a common potentially sight threatening ocular infection that may be caused by bacteria, fungi, viruses or parasites. Corneal ulcer is an ocular emergency that requires prompt management to ensure the best visual outcome for the patient. Otherwise, it can result in scarring and blindness. Aim and objectives: To study Microbial causes and to identify risk factors, and corelation of clinical features of ulcerative keratitis with etiological agents Materials and methods: Total 325 corneal ulcer cases were studied. Corneal scrapings were collected and subjected for examination by Gram's stain, 10% KOH, bacterial and fungal culture. Antibiotic susceptibility of the bacterial pathogens was performed by Kirby Bauer disc diffusion method as per CLSI guidelines with various groups of antibiotics. MIC for vancomycin were determined for study isolates by Epsilometer test. Results: Totally 325 infectious corneal ulcers were studied in detail. Aetiological agents were isolated in 115 (35.38%) cases. Majority of the isolates were fungal agents (44.18%), belonging to the Aspergillus (21.05%), Fusarium (14.47%). Bacterial corneal ulcer was less common (25.49%). The predominant bacterial pathogen isolated was Staphylococcus epidermidis, followed by Pseudomonas aeruginosa. no Acanthamoeba were identied. The sensitivity of potassium hydroxide staining was almost 100% in culture-proven fungal cases. Trauma is the leading cause for the corneal ulcers. Majority of the bacterial isolates were susceptible to amikacin. Conclusion: Ulcerative keratitis being a sight threatening disorder, early suspicion, rational use of laboratory diagnostic procedures, identication of the causative organisms and timely institution of appropriate antimicrobial therapy based on the prevailing sensitivity pattern of the isolates could save the eye from this preventable cause of blindness

Noninfectious posterior uveitis

Noninfectious posterior uveitis (NPU) comprises aheterogeneous group of vision-threatening, immune-mediated ocular and systemic diseases. It is predominantly bilateral and recurrent and, if not treated properly, leads to severe tissue damage that threatens the eyesight. In industrialized countries ca. 10-20% of all cases of blindness are caused by NPU. An NPU can occur at any age but is most common between the ages of20 and50 years. Laboratory diagnostic and imaging procedures enable an increasingly better differentiation of the disease spectrum. This makes it possible to better assess the course and prognosis of individual disease entities. An increasing repertoire of systemic and intravitreal forms of treatment has already led to more favorable long-term treatment outcomes. It can be expected that further progress can be achieved with better knowledge of the pathophysiology of the different clinical disorders and appropriate, targeted treatment.

Gynecological prerequisites for the development of chronic pelvic pain syndrome – diagnosis and treatment (Literature review)

Chronic pelvic pain syndrome is a complex process and mostly includes several organ systems. The gynecologic aspects of chronic pelvic pain syndrome can be divided into four distinct components: intra-abdominal pain, vaginal pain, pelvic floor pain, and sexual pain. The сommon gynecological causes of chronic pelvic pain are endometriosis, adenomyosis, vulvovaginal pain syndrome, pelvic floor muscle dysfunction, and sexual pain in women.This article describes the gynecological examination of patients with chronic pelvic pain and discusses the most common gynecological diseases and methods of their treatment. A review of the literature was conducted, which included the recommendations of the International Society for Urinary Incontinence, the European Association of Urology, and the International Association for the Study of Pain.Gynecological examination of patients with chronic pelvic pain begins with a history taking and physical examination. Laboratory tests, imaging examinations, and diagnostic procedures can be used additional methods to make a more accurate diagnosis. Treatment methods include physical therapy, medication, trigger point injections, and surgery.Because the diagnosis and treatment of chronic pelvic pain can be complex, it is important that physicians know the various aspects of this syndrome to be able to provide appropriate care for patients. Detailed history taking and physical examination for identifying the cause of the pain can help to determine the next step in evaluation and treatment. However, gynecological pathology is one of many, but not the only, that can be associated with chronic pelvic pain. This, it is necessary to perform a multimodal and multidisciplinary approach in the management of patients with chronic pelvic pain.

Clinical diagnostic techniques for rare genetic diseases in children: current status, advances, and thoughts

Rare diseases refer to a group of single diseases with low incidence rates, complex pathogeneses, severe disease conditions, and rapid progression. Most rare diseases have a genetic background and may occur in childhood. Paying attention to the rare genetic diseases in children and performing early diagnosis and treatment can effectively delay the course of disease and improve the quality of life of children. Many rare diseases can be diagnosed with the help of various experimental techniques, but the diagnosis of rare diseases is still not widely understood. This article summarizes the laboratory diagnostic techniques currently used for rare genetic diseases in children, so as to provide clues for the diagnosis and treatment of such diseases and help to enhance the theoretical understanding and precise medical treatment of rare genetic diseases in children.

Quantitatively detecting Candida albicans enolase1 with a one-step double monoclonal antibody sandwich ELISA assay.

Invasive candidiasis (IC) is often a cause of severe concern for the hospitalized patients, particularly those who are critically sick. However management of this disease is challenging due to a lack of effective laboratory diagnostic techniques. Hence, we have developed a one-step double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) using a pair of specific monoclonal antibodies (mAbs) for the quantitative detection of Candida albicans enolase1 (CaEno1), which is considered as an important diagnostic biomarker for IC. The diagnostic efficiency of the DAS-ELISA was evaluated by using a rabbit model of systemic candidiasis and compared with other assays. The method validation results demonstrated that the developed method was sensitive, reliable, and feasible. The findings of the rabbit model plasma analysis indicated that the diagnostic efficiency of the CaEno1 detection assay was better in comparison to the (1,3)-β-D-glucan detection and blood culture. CaEno1 is present in the blood of infected rabbits for a brief period and at relatively low levels and thus the combination of CaEno1 antigen and IgG antibodies detection could aid to increase diagnostic efficiency. However, to improve the clinical application of CaEno1 detection in the future, efforts should be made to increase the detection limit of the test by promoting technical developments and by optimizing the protocol for the clinical serial determinations.

Contemporary techniques to the laboratory diagnosis of cognitive dysfunction (Dementia) in elderly patients.

Dementia is a severe condition affecting more than 55 million people worldwide. In elderly patients, it leads to an overall decline in memory and thinking skills. It affects the patient and those around the patient, including family and caregivers. The symptoms can become so severe that the person's ability to perform everyday activities becomes compromised. This study aims to search and collect information about the sensitivity and specificity of available laboratory diagnostic methods for patients with Dementia and identify if these methods will be effective in giving a specific and definitive diagnosis. The specificity and sensitivity of four main laboratory testing methods have been included in this review: blood panels, cerebrospinal fluid, plasma biomarkers, hormone testing, and genetic testing. The methods are each explained separately with details of their importance and which specific parameters they screen for have haven included. Numerous blood panels and laboratory diagnostic techniques have been suggested for the timely and accurate diagnosis of Dementia and its various types; however, when the sensitivity and specificity of these techniques are considered, cerebrospinal fluid (CSF) and plasma biomarkers are found to be the most effective. Alzheimer's disease (AD) and other types of Dementia are now diagnosed using the biomarkers amyloid- (A), tau, and phosphorylated tau (p-tau181). With 93 percent accuracy in diagnosis, the innovative Precivity AD test created by the Washington University School of Medicine in St. Louis might be regarded as both highly sensitive and specific.

Anesthesia for thyroid surgery in heart transplant patients - first case study in Serbia

Introduction. The number of patients in the world who have undergone heart transplantation is increasing, with better and longer survival rates, and therefore the number of patients who undergo various surgical interventions after transplantation is also increasing. This presents a challenge for anaesthesiologists. Case outline. A 45-year-old female patient underwent a total thyroidectomy due to suspected thyroid cancer. She had a heart transplant three years earlier. Preoperative preparation included evaluation of cardiac function, consultative examinations by a transplant cardiologist, laboratory and other diagnostic procedures, as well as a detailed analysis of all 20 medications that the patient uses in daily therapy. Common drugs were used for premedication and general endotracheal anaesthesia, with careful dose titration. Medicines were also prepared for the occurrence of heart rhythm disorders, bearing in mind that the heart is denervated, but there was no need for their use. The operation and postoperative course went smoothly and on the third postoperative day the patient was discharged from the hospital in good general condition. Conclusion. Preoperative preparation, anaesthesia, and postoperative treatment of this patient represented a challenge for our team, which was successfully overcome, considering that this is the first case of operative treatment of a patient with a transplanted heart in Serbia.

Clinical insights and challenges: A rare case report of Leigh syndrome

Leigh syndrome is a severe neurological disorder that impairs mobility and respiration, among other bodily functions. Progressive brain deterioration is what makes it distinctive. This case report describes a 10-monthold female child who displayed symptoms such as vibratory movements in the right upper and left lower limbs, failure to thrive, decreased activity, and vomiting after consuming supplemental feeds. Multiple clinical signs of Leigh syndrome, including missed developmental milestones and mild acute malnutrition, were identified during a physical examination. So the symptoms that led to the diagnosis of Leigh syndrome were quadriparesis, dystonia, intermittent stridor (laryngeal dystonia), and bulbar weakness. This was supported by various laboratory results and diagnostic procedures, including brain imaging. The treatment included pacitane, thiamine, biotin, l-carnitine, calcium + vitamin d3, and sodium benzoate as well as dietary assistance and other medicines. Leigh syndrome is an incredibly rare disorder that has major effects on those affected and often manifests in infancy or early childhood. It is characterized by things like vomiting, feeding issues, neurological impairments, and delays in development. Sadly, because Leigh syndrome is a degenerative condition, the outlook for those who have it is frequently bleak. In cases with Leigh syndrome, this case emphasizes the value of early diagnosis and interdisciplinary treatment. The patient was instructed to have additional testing such as an EEG and a disability evaluation. Healthcare professionals must be aware of the clinical manifestation and treatment of Leigh syndrome in order to give affected people and their families the right care and support.

Decoding the Rare Diagnostic Enigma of Pythium Insidiosum Keratitis – A Narrative Review

Pythium insidiosum, an oomycete, is frequently referred to as a “parafungus” due to its striking similarity to fungal keratitis. Over the past 10 years, there has been a significant rise in Pythium insidiosum keratitis (PIK) cases, notably in Asia and India. This trend is potentially attributable to heightened research interest in this microorganism and advancements in diagnostic and therapeutic techniques. Clinically, its features, such as subepithelial infiltration, fluffy stromal infiltrates resembling cotton wool, satellite lesions, corneal perforation, endoexudates, and anterior chamber hypopyon, mirror those of fungal infections. However, hallmark clinical features like reticular dots, tentacular extensions, peripheral indentation, and prompt limbal progression distinguish Pythium from other pathogens and necessitate vigilant clinical scrutiny. From a morphological and microbiological perspective, Pythium closely mirrors fungi when subjected to routine smear tests, often revealing perpendicular or obtuse septate or non-septate branching hyphae. Culturing on nutritional media like blood agar, chocolate agar, and so on remains the diagnostic gold standard, with the organism presenting as cream-white colonies that form zoospores. This can be further verified through the leaf incarnation technique. Culture and polymerase chain reaction (PCR) are the gold standard diagnostic modalities for PIK. Historically, due to constraints in diagnostic techniques and the slow growth of cultures, there has been a trend toward employing advanced molecular diagnostic tools, including PCR, confocal microscopy, enzyme-linked immunosorbent assay, and immunodiffusion. When corneal scraping tests (10% KOH, Gram stain) depict fungal-like hyphae, antifungal treatments are often initiated even before culture results are ready. However, recent molecular research recommends the use of antibacterials, specifically 0.2% linezolid and 1% azithromycin, as the primary therapeutic agents. In cases that do not respond to treatment, early therapeutic keratoplasty is advised. This narrative review aims to provide a comprehensive overview of the epidemiology, clinical manifestations, laboratory and molecular diagnostic procedures, and therapeutic approaches for PIK.

Aspergillus and the Role of Cytopathology and Microbiology Lab Techniques in Diagnosis

Introduction: Aspergillosis is a type of fungal infection or a type of mold that can cause serious respiratory illnesses in people especially those with weakened immune systems. Timely diagnosis and treatment of Aspergillus infections is critical to preventing potentially life-threatening complications. It can be difficult to diagnose, as it often mimics other diseases. Cytopathology and microbiology lab techniques can be used to help diagnose aspergillosis; Histopathology, PCR and biochemistry are also important. Aims: To compare the most effective laboratory diagnostic techniques to detect the Aspergillus in body. Methods: This systematic review searched for studies that evaluated the use of cytopathology and microbiology and other lab techniques in the diagnosis of aspergillosis. Studies were identified through searches of Google Scholar, MEDLINE, EMBASE, and the Cochrane Library. Results: Cytopathology and microbiology lab techniques can both be helpful in diagnosing aspergillosis. Cytopathology and histopathology are both highly accurate, but results may take longer to obtain than with other tests. Microbiology is useful for diagnosing infections, but it may not be as accurate as cytopathology or histopathology. Poly Chain Reaction (PCR) is very accurate, but it is also expensive and results may take some time to obtain. Biochemistry can be useful for diagnosing metabolic disorders, but it is not as accurate as other tests. Conclusion: There is no one “best” way to detect Aspergillus, and the best approach may vary depending on the particular patient and situation. In general, a combination of two or more tests may be needed to accurately diagnose aspergillosis. The most important factor/guideline in choosing which tests to use is whether they will provide information that can help guide treatment decisions. Nevertheless, with all factors considered the PCR is the best test because it is highly sensitive and specific; able to detect even low levels of Aspergillus.

Cutaenous Leishmaniasis In Libya

Cutaneous Leishmaniasis is wide spread in northwestern of Libya with increasing of incidence in the last years, as well as with appearance of new foci in Tawarga, Sirte, and Zliten, to become also endemic areas, and added to old known endemic areas like Jabal Nafusa. Different clinical manifestations and atypical presentations of skin lesions necessitate more clinical alertness as well as use of laboratory diagnostic procedures. Leishmania major was the main cause of cutaneous Leishmaniasis in Libya, followed by L. tropica, which was less frequent cause in less than one-third of all molecularly investigated patients. Some of main used therapies in Libya were cryotherapy, and intralesional antimony for patients with few and small lesions, although, in some patients; with large number as well as large size of lesions or patients with appearance of lesions at face or ears; made systemic treatment with sodium stibogluconate or meglumine antimoniate more suitable and mandatory treatment. In Libya, low doses of antimony as intramuscular injection were the followed regime that was recommended by national guidelines. Oral rifampicin and isoniazid were also successful alternative systemic therapy when systemic antimony was unsuitable or not effective.

Hematological and serum biochemical profiles of a natural African swine fever virus infection in pigs.

African swine fever (ASF) is a contagious viral disease that affects pigs of all ages, inducing hemorrhagic fever with high mortality and severe threat to pig production. This study investigated the hematological and serum biochemical abnormalities associated with a natural ASF infection in pigs. A total of 100 serum samples of pigs from piggery suspected of ASFV infection were screened for antibodies by ELISA. Thirty‑two blood samples from serologically positive pigs and 32 negative pigs were undergo to hematological and serum biochemical analyses following standard procedures. The results showed that the mean values of the red blood cell (RBC) count, total white blood cell (TWBC) count, absolute lymphocyte count, absolute monocyte count, serum total protein (TP) and globulin were significantly (p < 0.05) lower while the mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), absolute neutrophil count and serum gamma glutamyl transferase (GGT) were significantly (p < 0.05) higher in the infected than the healthy pigs. There were no significant differences (p > 0.05) in the mean values of the packed cell volume (PCV), hemoglobin concentration, absolute eosinophil count, cholesterol, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities between the infected and healthy pigs. Hence, natural ASFV infection may have caused alterations in the hematological and serum biochemical parameters in the infected pigs. The generated data could complement the existing laboratory diagnostic techniques such as polymerase chain reaction, direct fluorescence antibody test, indirect fluorescent antibody test and ELISA in the diagnosis of ASF in pigs.

SOX1 antibody-related paraneoplastic neurological syndromes: clinical correlates and assessment of laboratory diagnostic techniques.

To describe the clinical associations of SOX1 antibodies (SOX1-Abs), determine the accuracy of various detection techniques, and propose laboratory criteria to identify definite paraneoplastic neurological syndromes (PNS) associated with SOX1-Abs. Single-center, retrospective study of patients referred to the French Reference Center between 2009 and 2019 for confirmation of SOX1-Ab positivity, without concurrent neural antibodies. Patients were classified according to the updated diagnostic PNS criteria; biological samples were systematically retested with three distinct techniques (line blot, cell-based assay, indirect immunofluorescence). Among 77 patients with isolated SOX1-Ab positivity, 23 (29.9%) fulfilled the criteria for definite PNS; all of them had lung cancer (mostly small-cell) and presented mainly with Lambert-Eaton myasthenic syndrome (10/23) and rapidly progressive cerebellar ataxia (6/23). SOX1-Ab positivity varied depending on the laboratory methods which were used, and a single technique was not sufficient to draw conclusions about the PNS diagnosis. The combination of an antigen-specific test (line blot and/or cell-based assay) and immunofluorescence showed the highest accuracy (81.5%, 95% CI 70.0-90.1) in identifying definite PNS. Moreover, when the PNS-Care score was recalculated assigning three points at the laboratory-level only to patients with positive "antigenic-specific test + immunofluorescence" and 0 points to the remaining cases, a higher certainty for definite and non-PNS was achieved (from 41/77, 53.2%, to 60/77, 77.9%; p < 0.001). SOX1-Abs should be considered high-risk antibodies only when detected with a positive antigenic-specific test and immunofluorescence. Other laboratory results and clinical associations different from Lambert-Eaton myasthenic syndrome and rapidly progressive cerebellar ataxia should be carefully reassessed to rule out false positivity and alternative diagnoses.

Potential Risk of Botulinum Neurotoxin -producing Clostridia Occurrence in Canned Fish.

Heat treatment is indispensable in fish canning to provide an acceptable shelf life. Its optimisation reduces the risk of the presence of Clostridium botulinum spores, which could potentially cause botulism cases. This study evaluated canned fish samples for botulism neurotoxin (BoNT)-producing clostridia contamination and can bulging through microbiological contaminant growth. A new analytical approach was developed for detection of such clostridia and phenotypically similar species. A total of 70 canned fish samples suspected of exhibiting bulging features were analysed. Culture methods were used to detect clostridia. The isolates obtained were evaluated on the basis of the exhibited phenotypic characteristics. Also, PCRs were used for the detection of genes determining BoNT production (non-toxic non-haemagglutinin (ntnh) genes) and the amplification of conservative 16S rDNA genes, which were Sanger sequenced. The obtained sequences were analysed using the Basic Local Alignment Search Tool. Clostridium genus species were isolated from 17 (24%) bulging and organoleptically changed samples. No ntnh genes were present in these isolates; however, sequencing confirmed the presence of C. sporogenes, a species with close affinity to C. botulinum. To eliminate the threat of foodborne botulism, laboratory diagnostic techniques must detect species of the Clostridium genus and elucidate their ability to produce BoNTs. Although Clostridium botulinum is the most common cause of botulism, the possibility may not be ignored that non-pathogenic Clostridium species may acquire botulinum toxigenicity. The similarity between the isolated strains of C. sporogenes and C. botulinum should be incorporated in the optimisation of heat treatment to guarantee a sterilised, microbiologically safe product.