Diagnostic Utility of Biplex/Multiplex Polymerase Chain Reaction in Infectious Granulomatous Dermatitis in North Indian Population.

BACKGROUND: A definite diagnosis of infectious granulomatous dermatitis (IGD) is difficult for both practicing dermatologists and dermatopathologists due to overlapping clinical and histomorphological features. We aimed to explore the role of multiplex polymerase chain reaction (PCR) for identifying a definite etiological agent for diagnosis and appropriate treatment in IGD in formalin-fixed paraffin-embedded tissue. MATERIALS AND METHODS: Sixty-two cases of IGD were included, excluding leprosy. The histochemical stains including Ziehl-Neelsen, periodic acid-Schiff, and Giemsa were performed in all cases. A multiplex PCR was designed for detection of tuberculosis (TB) (IS6110 and mpt64), fungal infections (ITS1, ITS2; ZM1, and ZM3), and leishmaniasis (kDNA). The results of histomorphology, histochemical stains, and multiplex PCR were compared. RESULTS: Among 62 cases, the sensitivity rate of PCR detection for organisms was 16.7%, 0%, 100%, 72%, 75%, and 66.7% in patients with TB, suggestive of TB, leishmaniasis, fungal infections, and granulomatous dermatitis not otherwise specified and granulomatous dermatitis suggestive of fungus, respectively. The TB PCR using IS6110 primers was negative in all cases; however, PCR using mpt64 primers was positive in 33.33% cases of scrofuloderma. The histochemical stains including Ziehl-Neelsen for acid-fast bacilli, periodic acid-Schiff for fungus, and Giemsa for Leishman-Donovan bodies showed positivity in 11.3%, 43.5%, and 3.2%, respectively. CONCLUSION: A multiplex PCR (Mycobacterium tuberculosis, Leishmania, and panfungal) is highly recommended in all cases of IGD where an etiological agent is difficult to establish by skin biopsy and histochemical stains along with a clinicopathological correlation. This will augment in appropriate treatment and will reduce empirical treatment and morbidity in such patients.

Seek and Find! PCR analyses of skin infections in West-European travelers returning from abroad with an eschar.

BACKGROUND: Skin infections are among the leading causes of diseases in travelers. Diagnosing pathogens could be difficult. METHOD: We applied molecular assays for the diagnostic of a large collection of skin biopsies and swabs from travelers with suspected skin infections. All samples were tested by qPCR for Coxiella burnetti, Bartonella sp., Rickettsia sp., Borrelia sp., Ehrlichia sp., Tropheryma whipplei, Francisella tularensis, Mycobacteria sp., Staphylococcus aureus, Streptococcus pyogenes, Leishmania spp., Ortho poxvirus and Para poxvirus and then screened for the presence of bacteria by PCR amplification and sequencing, targeting the 16S rRNA gene. RESULTS: From January 2009 to January 2017, 100 international travelers presenting with a suspected skin infection were enrolled. We detected 51 patients with an identified pathogen on skin samples. Travelers presenting with eschars were more likely to have a positive PCR sample (n = 44/76, 57.9%) compared to other patients (n = 7/24, 29.2%). Spotted fever group Rickettsia (n = 28) was the most frequently detected pathogens (19 R. africae, 6 R. conorii, 3 R. mongolitimonae); S. aureus were detected in 11 patients; S. pyogenes in 3; Leishmania sp.; M. leprae and B. henselae in 1 patient, respectively. CONCLUSION: By targeting the most commonly encountered causative agents of travel-related skin infections, our strategy provides a sensitive and rapid diagnostic method.

Tropical skin diseases in British military personnel.

Skin complaints are common in travellers to foreign countries and are responsible for up to 25% of medical consultations by military personnel during deployments in the tropics. They also have relatively high rates of field hospital admission, medical evacuation and referral to UK Role 4 healthcare facilities. Non-infectious tropical skin diseases include sunburn, heat rash, arthropod bites, venomous bites, contact dermatitis and phytophotodermatitis. During tropical deployments skin infections that commonly occur in military personnel may become more frequent, severe and difficult to treat. Several systemic tropical infections have cutaneous features that can be useful in making early diagnoses. Tropical skin infections such as cutaneous larva migrans, cutaneous myiasis, cutaneous leishmaniasis and leprosy do occur in British troops and require specialist clinical management. This illustrated review focuses on the most significant tropical skin diseases that have occurred in British military personnel in recent years. Clinical management of these conditions on deployments would be improved and medical evacuations could be reduced if a military dermatology 'reach-back' service (including a telemedicine facility) was available.

Infection and disease caused by environmental mycobacteria.

Many species of mycobacteria that normally live as environmental saprophytes, the environmental mycobacteria (EM), are opportunist causes of disease in humans and animals. Many, but not all, cases are associated with some form of immune deficiency. An increasing number of species and clinical presentations are being described, and advances are being made in the understanding of the underlying predisposing factors. In recent years, four aspects of EM disease have become particularly relevant to human health: (1) the high prevalence of EM disease in patients with AIDS; (2) the emergence of Buruli ulcer, an ulcerative skin disease caused by Mycobacterium ulcerans, as the third most prevalent mycobacterial disease; (3) the effect of infection by EM on the immune responses to BCG vaccination and on the course and outcome of tuberculosis and leprosy; (4) the controversy over the involvement of mycobacteria, notably M. avium subspecies paratuberculosis, in human inflammatory bowel disease. These aspects change the status of EM from mere curiosities to important direct, indirect, and putative causes of serious and increasingly common human disease.

Non-cultivable mycobacteria in ulcers of the skin.

In biopsies of 54 patients suffering from chronic dermatological lesions (mostly ulcers of the skin) acid-fast bacilli were found in 14. In these 14 cases in 4 were lesions caused by M. tuberculosis, in 1 the lesion was caused by M. avium-intracellulare, in 1 the lesion was caused by M. fortuitum and in 2 the lesions were caused by non-cultivable mycobacteria (Feldmann-Hershfield ulcer?). In 2 cases the cultures were heavily contaminated, and the diagnosis remained uncertain. In the remaining 4 cases the mycobacteria were considered occasional isolates without clinical significance.

The pathology of Mycobacterium ulcerans infection.

The pathology of cutaneous ulcers resulting from Mycobacterium ulcerans infection is reviewed. Initial infection causes ulceration with necrosis of the dermis and a septate panniculitis in subcutaneous fat. There is little cellular reaction despite the presence of large numbers of organisms. Recurrent or persistent infection produces a granulomatous reaction with epithelioid macrophages, variable numbers of giant cells of the Langhans type, and relatively few organisms. This type of reaction is associated with more successful treatment of the disease and appears analogous to the tuberculoid form of leprosy.

The cutaneous infiltrates of leprosy. A transmission electron microscopy study.

The dermal lesions of 18 patients with leprosy have been examined by transmission electron microscopy. The patients exhibited a spectrum of disease from polar lepromatous to polar tuberculoid with intermediate stages in various states of therapy and relapse. The nature and quantities of inflammatory cells and bacteria have been determined by electron microscopy to supplement previous light and fluorescence microscopy studies. Lepromatous leprosy was characterized by many parasitized foam cells containing large, multibacillary vacuoles with intact, osmiophilic Mycobacterium leprae: Bacteria were embedded in an electron-lucent matrix. No extracellular bacteria were evident. Only small numbers of scattered lymphocytes were found. As one approached the borderline state, smaller numbers of bacilli were present as singlets and doublets in small vacuoles of macrophages. The more reactive forms showed increasing bacillary fragmentation, larger numbers of lymphoid cells, and an occasional epithelioid cell. At the tuberculoid end of the spectrum, clear evidence of an exuberant lymphocyte response was evident. Large numbers of T cells with extremely long and complex filipodia were closely associated with epithelioid and multinucleated giant cells. Many of the mononuclear phagocytes appeared nonviable, and areas of necrosis were evident. Bacillary remnants were scarce and the cytoplasm of the epithelioid cells contained occasional dense bodies and many stacks of endoplasmic reticulum and mitochondria. These results suggest that Leu 3a/OKT4 helper cells may be capable of driving the effector function of mononuclear phagocytes. This would lead to a significant microbicidal effect on M. leprae, perhaps through the production of toxic oxygen intermediates.