Cytokine profile of patients with leptospirosis in Sabah, Malaysia

Background: Leptospirosis is a zoonotic disease withsymptoms ranging from a mild, febrile illness to a severeform with multiorgan failure. Severe leptospirosis mayrequire medical interventions in the form of dialysis and/ormechanical ventilation and often leads to mortality. Anexaggerated host immune response—in particular, a“cytokine storm”—that causes endothelial and organdamage is associated with the disease severity andmortality.Methods: Microscopic agglutination test (MAT)-positive andMAT-negative human serum samples (n=30) from patientswith leptospirosis were obtained from the Public HealthLaboratory, Kota Kinabalu, Sabah, Malaysia and controlserum samples (n=10) were obtained from healthy studentvolunteers. We estimated the levels of IL-1β, IL-6, IL-8, IL-10,and TNF-α in serum samples by a Luminex assay.Results: The levels of IL-6, IL-8, and IL1-β were significantlyhigher in 13% of the patients with leptospirosis compared tothe healthy controls, while the levels of IL-10 and TNF-α werenot elevated in either group.Conclusion: Our data suggest that elevated levels of IL-6, IL-8, and IL1-β may be associated with leptospirosis diseaseseverity, which requires patient follow-up for confirmation.

Recombinant LipL32 Protein Developed Using a Synthetic Gene Detects Leptospiraspecific Antibodies in Human Serum Samples

Background: Synthetic biology is emerging as a viable alternative for the production of recombinant antigens for diagnostic applications. It offers a safe alternative for the synthesis of antigenic principles derived from organisms that pose a high biological risk. Methods: Here, we describe an enzyme-linked immunosorbent assay (ELISA) using the synthetic recombinant LipL32 (rLipL32) protein expressed in Escherichia coli for the detection of Leptospira-specific antibodies in human serum samples. The rLipL32-based ELISA was compared with a microscopic agglutination test (MAT), which is currently used as the gold standard for the diagnosis of leptospirosis. Results: Our results showed that all the MAT-positive serum samples were positive for Leptospira-specific IgG in an ELISA, while 65% (n = 13) of these samples were also positive for Leptospira-specific IgM. In the MAT-negative serum samples, 80% and 55% of the samples were detected as negative by an ELISA for Leptospira-specific IgM and IgG, respectively. Conclusion: An ELISA using the synthetic rLipL32 antigen was able to distinguish Leptospira-specific IgM (sensitivity 65% and specificity 80%) and IgG (sensitivity 100% and specificity 55%) in human serum samples and has the potential to serve as a rapid diagnostic test for leptospirosis.

Characteristics, Detection And Typing Methods Of Community Acquired Methicillin Resistant Staphylococcus Aureus (CA-MRSA) – A Review

@#Staphylococcus aureus aregram positive cocci which colonizethe skin and mucous membranes particularly the anterior nares. Prevalence of nosocomial infections associated with methicillin resistant S. aureus have been reported in hospitals (HA-MRSA) for over five decades. Recently,community-acquired MRSA (CA-MRSA) has emerged as a cause of skin and soft tissue infections in healthy individuals. These strains are sensitive to antimicrobials, carry genes for Panton-Valentine leukocidin (PVL) toxin and belong to the staphylococcal cassette chromosome (SCC) mec type IV or V. The suspected mode of transmission involves close contact with carriers leading to skin or nasal colonization that resultin subsequent active infection. Molecular typing is used to determine the mode of transmission of CA-MRSA in the community.General typing methods such as pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) and specific methods for Staphylococci such as SCCmec typing and spa typing have the capability to characterize bacterial chromosomes and mobile genetic elements. Combination of these molecular typing methods is necessary as each method has its own advantages with respect to discriminatory power, rapidity, cost effectiveness, reproducibility, and ease of performance.

Leptospirosis: recent incidents and available diagnostics – a review

Objective: The aim of this article was to review published research articles on leptospirosis, in particular the recent incidence of leptospirosis in Malaysia and the currently available diagnostic methods for the detection of leptospirosis. Methods: PubMed, Google Scholar and Google Search databases were searched using the key words Leptospira and leptospirosis. A total of seventy-six references were reviewed including sixty-seven research articles, three annual reports from Ministry of Health and six online newspaper articles. This review includes the following five sub-headings: introduction, leptospirosis transmission, leptospirosis incidents, laboratory diagnosis of leptospirosis and treatment and prevention of leptospirosis. Results: An increase in incidents of leptospirosis cases has been seen in recent years in Malaysia. The recent floods have contributed to the rise in the number of reported cases. Current diagnostic approaches such as dark field microscopy, microscopic agglutination test (MAT), Polymerase chain reaction and serological tests are inadequate as the organism is a slow grower. Conclusion: There is an urgent need to develop newer techniques for rapid detection of leptospirosis. The combination of PCR and ELISA are suggested for rapid and accurate diagnosis of leptospirosis. Studies on the mechanism of pathogenesis of Leptospira are needed for the development of vaccines that are safe for human use.

Diagnosis of tuberculous meningitis: challenges and promises

Tuberculosis (TB) which is caused by Mycobacterium tuberculosis infects primarily the lungs but it also affects other parts of the body. Tuberculous meningitis (TBM) is the most severe form of TB and has the highest mortality and morbidity rate compared to other forms of TB. It is common in young children and HIV-infected patients, but is also seen in adults. Despite anti-tuberculosis treatment, TBM is still a major cause of death and neurological sequelae as treatment given to the patients is often delayed. Early diagnosis is challenging due to the non-specific symptoms of TBM and the low number of tubercle bacilli in cerebrospinal fluid (CSF). Until now, there is no established diagnostic method that can rapidly detect M. tuberculosis in TBM patients with high sensitivity and specificity. The emergence of drug resistant M. tuberculosis strains further complicates the diagnosis and treatment regimen of TBM. This review summarizes challenges of the currently used diagnostic methods and the potential future use of molecular diagnostic methods for TBM.