First case of Paragonimus westermani infection in a female patient in India.

Paragonimiasis is a foodborne parasitic zoonosis caused by lung fl uke species of the genus Paragonimus. The Paragonimus westermani is the most common human pathogen in Asian countries. In northeast India, Paragonimus heterotremus has been documented as the only human pathogen in the earlier literature. In India, P. westermani infection in humans remained undetermined. Herein, we report a case of pulmonary paragonimiasis due to P. westermani in an adult female in Manipur. The diagnosis was made by morphological and molecular characterisation of the eggs in the sputum. This is the fi rst confi rmed case of paragonimiasis due to P. westermani in India.

Subcutaneous nocardial abscess in a post-renal transplant patient.

A case of nocardiosis in a post-renal transplant patient is being reported for the first time from Eastern India. The patient had multiple abscesses on the right thigh and chest. Direct examination of the aspirated pus by Gram stain and modified Ziehl Neelsen stain revealed gram-positive beaded, partially acid-fast, branching filaments. Culture of the pus yielded Nocardia asteroides. The patient responded to surgical drainage and sulphamethoxazole therapy.

Emergence of sporotrichosis in Manipur.

A retrospective analysis of all cases of sporotrichosis was conducted to examine the pattern and frequency of sporotrichosis cases in Manipur over a period of six year from July 1999 to June 2005. Seventy-three (73) cases of sporotrichosis were detected of which 30 were confirmed by culture and 43 were diagnosed by aspiration cytology only. Most of the patients belonged to the age group of 21 to 40 years (n = 23; 31.5%). There were 39 females (53.4%) and 34 males (46.5%). Commonest site of infection was upper limbs (n = 39; 53.4%) followed by lower limbs (n = 17; 23.2%). Commonest type of infection was lymphocutaneous form (n = 46; 63.1%) followed by fixed cutaneous form (n = 27; 36.9%). Among these 73 cases, 2 male patients (2.7%) were found to be HIV positive. In our experience, collection of material by aspiration of pus or infected tissue was found to be a better method compared to scraping or exudate. This study revealed Manipur as a new endemic area for sporotrichosis.

Isolation and evaluation of diagnostic value of two major secreted proteins of Mycobacterium tuberculosis.

Two secreted antigens of Mycobacterium tuberculosis, namely the antigen 85 complex (30/31) and 38kDa antigens, were purified from the whole culture filtrate by using two dimensional preparative electrophoresis and anion exchange chromatography, respectively. Individual components of the antigen 85 complex namely, antigen 85A, 85B and 85C, were separated using hydrophobic interaction chromatography. The humoral antibody activity to these antigens in sputum positive cases of active pulmonary tuberculosis and normal healthy volunteers was determined by enzyme linked immunosorbent assay (ELISA) and immunoblot. Recombinant 38kDa and antigen 6 were used as reference antigens for the assay. None of the healthy volunteers reacted with the 38kDa antigen, while 52% of the TB sera reacted with it. Of the three components of the antigen 85 complex, 85B gave the highest positivity of 40 per cent. The results of combination of 38kDa with antigen 6 offered better results with 76% positivity.

Production & characterization of monoclonal antibodies to Mycobacterium tuberculosis.

BACKGROUND & OBJECTIVES: Monoclonal antibodies (MAbs) against Mycobacterium tuberculosis H37Rv culture filtrate (CF) were raised by immunizing BALB/c mice and characterization was done. Attempts have been directed towards identifying mycobacterial antigens in biological fluids by employing polyclonal and monoclonal antibodies specific for M. tuberculosis. Immunohistologic studies, using MAbs for the localization of whole or fragmented bacilli in the biopsy specimens were also carried out. METHODS: Intrasplenic IS and intraperitoneal i.p. routes of immunization, were compared. The MAbs were characterized for their isotype, binding specificity, nature of binding epitope, reactivity in immunoassays etc. RESULTS: IS and i.p. routes of immunization, were compared and i.p. was found superior. Ten MAbs designated TRC 1-10 were produced. Of these, 7 MAbs, TRC 1-7 reacted with the 30/31 kDa doublet (antigen 85 complex), TRC 8 with 12 kDa in addition to 30/31 kDa and TRC 9 and 10 with the 24 and 12 kDa antigens respectively. Six MAbs were classified as broadly cross reactive and 2 showed limited cross reactivity. TRC 8 and 10 showed species specificity. Employing TRC 8 in sandwich ELISA, antigen was detected in sera from 17 of 25 pulmonary tuberculosis patients and 3 of 20 controls. TRC 8 was found to be useful in detecting antigens specifically in M. tuberculosis and M. leprae infected tissues, by immunoperoxidase staining. INTERPRETATION & CONCLUSION: TRC 8 was found to be restricted in its reactivity to M. tuberculosis complex and M. leprae. TRC 8 may prove useful in immuno-diagnosis of tuberculosis.