Mycobacterial Polymerase Chain Reaction Research Articles

Hepatic microgranulomas caused by Mycobacterium marinum (MM) in a Horsfield tortoise (Testudo horsfieldii) presenting with a non‐healing limb granuloma

AbstractA 3‐year‐old, male, entire Horsfield tortoise (Testudo horsfieldii) presented with a non‐healing lesion on the dorsal aspect of the elbow. Fine‐needle aspiration cytology revealed granulomatous inflammation with acid‐fast staining organisms. Due to worsening clinical condition, the animal was euthanased. Postmortem examination included histopathology, Mycobacterium culture and polymerase chain reaction testing. There was gross hepatomegaly and a large, turbid, straw‐coloured, coelomic effusion. Limb granuloma and liver tissue were sent for mycobacterial polymerase chain reaction and culture, and both were positive and confirmed disseminated Mycobacteria marinum. The origin of the infection is not known, but suspected to originate from poor hygiene from the place of purchase and dissemination secondary to the initial limb granuloma. This represents an uncommon presentation and the first reported infection by this mycobacterial agent in the species. It is important to consider mycobacterial disease as a differential in granulomatous infections, given the range of presentations possible in reptile patients.

Granulomatous Rosacea Versus Lupus Miliaris Disseminatus Faciei-2 Faces of Facial Granulomatous Disorder: A Clinicohistological and Molecular Study.

Granulomatous rosacea (GR) and lupus miliaris disseminatus faciei (LMDF) are 2 forms of facial granulomatous diseases. Although they show some morphological overlap, they have distinct clinical presentation. This study was performed to demonstrate the clinical and histological features of GR and LMDF and to establish their relationship to tuberculous etiology by molecular technique. All the cases of GR (n = 20) and LMDF (n = 10) diagnosed on skin biopsy over the past 6 years were reviewed along with their clinical detail. Polymerase chain reaction (PCR) was performed using primers specific for Mycobacterium tuberculosis. The mean age of patients with GR was 45 years 10 months (range 18-75 years) as compared to 33 years 5 months (range 18-57 years) in patients with LMDF. The GR cases comprised 13 men and 7 women patients, whereas all 10 LMDF cases were seen in men. GR cases had papular lesion over an erythematous base on face, whereas LMDF cases had papular/nodular/nodulocystic lesions on the face and neck. Histologically, GR cases showed small granulomas without necrosis in a background of variable lymphoid infiltrate and dilated capillaries, whereas LMDF showed large granulomas with caseous necrosis and minimal inflammation. Five cases (25%) of GR showed degenerating Demodex folliculorum mites. No case of GR or LMDF showed positivity for mycobacterial polymerase chain reaction. Despite some similarities, GR and LMDF show distinct clinical and histological features. Thus, LMDF is a distinct clinicopathological entity separate from the GR, with different etiopathogenesis. However, none of the conditions are related to a tuberculous etiology.

Clinical and Pathological Evaluation of Mycobacterium marinum Group Skin Infections Associated With Fish Markets in New York City.

From December 2013 through May 2014, physicians, dermatopathologists, and public health authorities collaborated to characterize an outbreak of Mycobacterium marinum and other nontuberculous mycobacterial skin and soft tissue infections (SSTIs) associated with handling fish in New York City's Chinatown. Clinicopathologic and laboratory investigations were performed on a series of patients. Medical records were reviewed for 29 patients. Culture results were available for 27 patients and 24 biopsy specimens were evaluated by histopathology, immunohistochemistry (IHC) staining for acid-fast bacilli (AFB), and mycobacterial polymerase chain reaction (PCR) assays. All patients received antibiotics. The most commonly prescribed antibiotic regimen was clarithromycin and ethambutol. Of the 29 patients in this case series, 16 (55%) received surgical treatment involving incision and drainage, mass excision, and synovectomy. Of these, 7 (44%) had deep tissue involvement. All patients showed improvement. For those with culture results, 11 of 27 (41%) were positive for M. marinum; the remainder showed no growth. Poorly formed granulomas (96%), neutrophils (75%), and necrosis (79%) were found in 24 biopsies. Of 15 cases that were culture-negative and analyzed by other methods, 9 were PCR positive for M. marinum group species, 8 were IHC positive, and 3 were positive by AFB stains. A multidisciplinary approach was used to identify cases in an outbreak of M. marinum infections. The use of histopathology, culture, and IHC plus PCR from full thickness skin biopsy can lead to improved diagnosis of M. marinum SSTIs compared to relying solely on mycobacterial culture, the current gold standard.

Sequelae of World War II: An Outbreak of Chronic Cutaneous Nontuberculous Mycobacterial Infection among Satowanese Islanders

After World War II, residents of Satowan (population, 650 persons), an outer island in the state of Chuuk, Federated States of Micronesia, noted a high prevalence of a chronic, progressive skin disease known locally as "spam." Island residents who had chronic, progressive verrucous or keloidal plaques for >3 months were considered case patients. Tissue specimens were obtained for culture, histopathological analysis, mycobacterial polymerase chain reaction (PCR), and comparison with the hsp65 gene of Mycobacterium marinum. We performed a case-control study involving all cases and randomly selected control individuals from the community. A total of 39 case patients were identified, with a median age of 26.0 years (range, 8-82 years); 74.4% were male, and the mean duration of disease was 12.5 years. A total of 98 control individuals were enrolled. Results of all 19 mycobacterial tissue cultures were negative, and histopathological analysis of all 9 lesions showed suppurative granulomatous inflammation with negative results of mycobacterial and fungal stains. In 7 of 9 paraffin-embedded samples, nontuberculous mycobacterial DNA was detected by PCR, and 2 sequenced products had 95% and 87% identity to M. marinum. All case patients were taro farmers (odds ratio, undefined; P < .01), and among taro farmers, when the analysis was controlled for sex, contact with water-filled World War II-era bomb craters was associated with infection (odds ratio, 8.2; P < .01). "Spam disease" is a chronic, progressive skin disease of high prevalence on Satowan and is associated with taro farming and contact with World War II-era bomb craters. Histopathological and PCR data demonstrate a nontuberculous mycobacterial infection as the cause.

Kidney Biopsy Findings in a Patient With Fever, Bilateral Pulmonary Infiltrates, and Acute Renal Failure

Kidney Biopsy Findings in a Patient With Fever, Bilateral Pulmonary Infiltrates, and Acute Renal Failure

A search for mycobacterial DNA in sarcoidosis using the polymerase chain reaction.

The etiology of sarcoidosis is unknown, but mycobacteria have been considered as a possible etiologic agent. The authors used the polymerase chain reaction (PCR) to search for mycobacterial DNA in paraffin-embedded granulomatous tissues from patients with sarcoidosis. The target sequence used for PCR amplification is a 383-base pair segment of the gene encoding the 65 kD mycobacterial surface antigen. This assay can detect Mycobacterium tuberculosis and atypical mycobacteria in archival material. Its sensitivity, which is superior to Ziehl-Nielsen staining for acid-fast bacilli, is 1 bacterium per 2500 cells. Ten sarcoidosis blocks and 10 normal controls were negative with mycobacterial PCR but positive with beta-actin PCR, indicating the presence of amplifiable DNA. Mycobacterial PCR gave positive results for six acid-fast bacilli stain/culture-positive blocks from patients with tuberculosis. These results indicate that sarcoidosis probably does not represent an active mycobacterial infection. These data also suggest that mycobacterial PCR is helpful in differentiating tuberculosis and sarcoidosis.

Identification of spheroplast-like agents isolated from tissues of patients with Crohn's disease and control tissues by polymerase chain reaction.

Mycobacterium paratuberculosis has been isolated from tissue taken from patients with Crohn's disease and has been implicated in the etiology of this disease. On culture, the organisms appear initially as cell wall-deficient, spheroplast-like forms that are difficult to identify by conventional techniques. Here we examine 30 unidentified cultures by the polymerase chain reaction using primers specific for M. paratuberculosis, Mycobacterium tuberculosis, and Mycobacterium avium restriction fragment length polymorphism type A/I and also by a non-species-specific mycobacterial polymerase chain reaction. Six of these cultures, all from Crohn's disease, were shown to contain DNA from M. paratuberculosis. Cultures from both Crohn's disease and controls were found to contain mycobacterial DNA of unknown specific origin.