Rapid diagnosis of Leishmania infection with a portable loopmediated isothermal amplification device.

L. donovani is an intracellular protozoan parasite, that causes visceral leishmaniasis (VL), and consequently, post-kala azar dermal leishmaniasis (PKDL). Diagnosis and treatment of leishmaniasis is crucial for decreasing its transmission. Various diagnostic techniques like microscopy, enzyme-linked immunosorbent assays (ELISA) and PCR-based methods are used to detect leishmaniasis infection. More recently, loop-mediated isothermal amplification (LAMP) assay has emerged as an ideal diagnostic measure for leishmaniasis, primarily due to its accuracy, speed and simplicity. However, point-of-care diagnosis is still not been tested with the LAMP assay. We have developed a portable LAMP device for the monitoring of Leishmania infection. The LAMP assay performed using our device can detect and amplify as little as 100 femtograms of L. donovani DNA. In a preliminary study, we have shown that the device can also amplify L. donovani DNA present in VL and PKDL patient samples with high sensitivity (100%), specificity (98%) and accuracy (99%), and can be used both for diagnostic and prognostic analysis. To our knowledge, this is the first report to describe the development and application of a portable LAMP device which has the potential to evolve as a point-of-care diagnostic and prognostic tool for Leishmania infections in future.

de novo Histoid leprosy: an expatriate case recently diagnosed in Johannesburg

Abstract: Histoid leprosy (HL) is a rare variant of lepromatous leprosy with unique clinical, histopathological, and microbiological features. A 32-year-old man from Malawi who immigrated to Johannesburg 1-year-ago, presented with a 4-month history of flesh-colored nodules on the face and trunk and hyperpigmented plaques on the chest and limbs. Skin slit smears confirmed multibacillary leprosy, and skin punch biopsies showed proliferation of spindled cells containing a large number of acid-fast bacilli. The prevalence of de novo HL is increasing in the era of leprosy elimination. HL cases may act as reservoirs and negatively affect the global control of leprosy.

de novo Histoid leprosy: an expatriate case recently diagnosed in Johannesburg.

Histoid leprosy (HL) is a rare variant of lepromatous leprosy with unique clinical, histopathological, and microbiological features. A 32-year-old man from Malawi who immigrated to Johannesburg 1-year-ago, presented with a 4-month history of flesh-colored nodules on the face and trunk and hyperpigmented plaques on the chest and limbs. Skin slit smears confirmed multibacillary leprosy, and skin punch biopsies showed proliferation of spindled cells containing a large number of acid-fast bacilli. The prevalence of de novo HL is increasing in the era of leprosy elimination. HL cases may act as reservoirs and negatively affect the global control of leprosy.

Correlates of GLA family adjuvants' activities.

Lipopolysaccharide (LPS) is a well-defined agonist of Toll-like receptor (TLR) 4 that activates innate immune responses and influences the development of the adaptive response during infection with Gram-negative bacteria. Many years ago, Dr. Edgar Ribi separated the adjuvant activity of LPS from its toxic effects, an effort that led to the development of monophosphoryl lipid A (MPL). MPL, derived from Salmonella minnesota R595, has progressed through clinical development and is now used in various product-enabling formulations to support the generation of antigen-specific responses in several commercial and preclinical vaccines. We have generated several synthetic lipid A molecules, foremost glucopyranosyl lipid adjuvant (GLA) and second-generation lipid adjuvant (SLA), and have advanced these to clinical trial for various indications. In this review we summarize the potential and current positioning of TLR4-based adjuvant formulations in approved and emerging vaccines.

Scabies Among Elderly Korean Patients with Histories of Leprosy.

A scabies epidemic, traced by the hospital-based surveillance system, was reported in a Korean leprosarium. A total of 200 symptomatic cases were found during 2012-2014 among 570 elderly former leprosy patients. Most of cases were classic type scabies (87%) and aged 75 years and older (72%). Surveillance system for early diagnosis and prompt intervention was applied and the scabies epidemic was controlled effectively in this long-term care facility.

Characterization of microbial flora of leprous ulcers infested with maggots.

Swabs from 64 maggot infested leprosy ulcers before and after treatment for maggots and 100 non-infested leprosy ulcers were studied for the bacterial flora. From maggot infested ulcers (before treatment), the cultures usually showed mixed growth. Among the Gram positive isolates, Staphylococcus aureus (37%), S. albus (18%) and Streptococcus pyogenes (36%) were frequently isolated. Gram negative bacteria isolated were Proteus spp. (21%) and Escherichia coli (7%). Anaerobic bacteria isolated were Micrococcus (3%) and Bacteroides (4%). After treatment of maggot infested ulcers, S. aureus (36%) continued to be isolated with almost the same frequency. The isolates of other Gram negative organisms were slightly reduced. Among the Gram negatives the Proteus spp. (10%) were also less in number. In few cases Neisseria (3%) was found. Anaerobic isolates were M. luteus (2%) and B. necrophorus (3%). From the cases without maggot infestation, a single organism was isolated from 16 cases and 84 mixed cultures were obtained. Isolates included the aerobic Gram positives S. aureus (46%), S. albus (21%) and S. pyogenes (38%), and the Gram negative Proteus spp. (19%) and E. coli (7%). The anaerobic isolate was M. luteus (3%). From this study no apparent association between the type of bacterial flora and maggot infestation could be observed.

Sistema monocitico-macrofagico y Mycobacterium leprae: su importancia en la interpretacion patogenica de las distintas formas clinico-patologicas de la lepra y de las reacciones leprosas / Monocyte-macrophage system and Mycobacterium leprae: its importance in the patogen interpretation of the different clinic-patology forms of the leprosy and the leprous reactions

Se realiza una revision de la patogenia de la lepra (Hanseniasis), en sus vinculaciones con el "sistema monotitico-macrofagico" y se efectua el estudio histopatologico, histoquimico (lipidico) y con microscopia electronica de cuatro enfermos de lepra, dos lepromatosos (sin y con eritema nudoso) y otros dos dimorfos ("Borderline"). Se concluye que: 1) La lepra presenta sus manifestaciones clinico-patologicas mas ostensibles, en vinculacion con lesiones del "sistema manocitico-macrofagico". 2) Existem distintos comportamientos de los macrofagos frente al Mycobacterium leprae: a) en personas Mitsuda-positivos de puede demostrar el predominio de "macrofagos lisadores del M. leprae" con muerte y desaparicion de los bacilos; b)en individuos Mitsuda-negativos predominan los "macrofagos no-lisadores del M. leprae", que tambiem producen al muerte de los bacilos, pero con persistencia de la "envoltura lipidica bacilar", que se deposita en las vacuolas de los virchowianos: y c) en enfermos Mitsuda-negativos, luego de fromado el "granuloma virchowiano", se desarrollan los "macrofagos lisadores de las celulas de Virchow" que permitiram el recambio celular de los virchowianos. 3) los "macrofagos lisadores del M. leprae" obtienen informacion antigenica del "bacilo aislado" que puedem transmitir al sistema de inmunidad celular, cuyos linfocitos T activados generan los "granulomas epitelioides con celulas de Langhans" (granulomas tuberculoides), de la Lepra tuberculoide. 4) Los "macrofagos no-lisadores del M. leprae" no obtendrian infromacion antigenica adecuada y no tendrian capacidad para estimular ningun sistema inmunologico (celular, o humoral), originando los "granulomas virchowianos" de la lepra lepromatosa. 5) Luego del envejecimiento y muerte de los virchowianos, los "macrofagos lisadores de las celulas de Virchow" obtendrian informacion antigenica de los "bacilos degenerados asociados a lipidos y glucoproteinas citoplasmaticas", con capacidad de estimulacion del sistema inmunologico humoral cuyos linfocitos B activados generarian anticuerpos complejos: a) contra bacilos ubicados en los virchowianos en la reaccion leprosa tipo 2 (eritema nudoso leprotico); b) contra la pared de vasos en la reaccion leprosa tipo 3 (fenomeno de Lucio); y c) contra tejidos del huesped (enfermedad autoagresiva hanseniana-Azulay)