Role of PCR for diagnosing Pneumocystis jirovecii pneumonia in HIV-infected individuals in a tertiary care hospital in India.

Objectives : In developing countries like India, the diagnosis of Pneumocystis jirovecii infection is often made either by conventional staining or clinically. This study was planned to know the utility of polymerase chain reaction (PCR) in diagnosing Pneumocystis jirovecii pneumonia (PJP) in human immunodeficiency virus (HIV)-infected patients, to compare the PCR results with that of staining techniques and also to correlate the results with clinical condition of patients. Materials and Methods: A prospective study included 50 HIV-infected adult in-patients with symptoms of lower respiratory tract infection. Induced sputum, bronchoalveolar lavage or tracheal aspirate were proceeded for both staining and PCR for mitochondrial large subunit rRNA gene of P. jirovecii. Results: In our study PCR results correlated with staining findings in 14% (7/50) of cases. Another 20% (10/50) cases could be diagnosed only with PCR, where staining was negative for the presence of P. jirovecii. When compared with clinical evidence of disease, PCR showed 93.7% sensitivity and 94.1% specificity. Presence of dyspnea and CD 4 count showed statistical significance (P<0.05) in PCP-diagnosed patients. Conclusions: PCR can be used for early and accurate diagnosis of PCP in HIV-infected patients.

Comparação entre três métodos de coloração a frio no diagnóstico primário de tuberculose: um estudo piloto / Comparison among three cold staining methods in the primary diagnosis of tuberculosis: a pilot study

OBJETIVO: Em países em desenvolvimento, a baciloscopia é a principal ferramenta para a identificação de casos de tuberculose pulmonar. O objetivo do presente estudo foi avaliar a eficácia diagnóstica do método de coloração de Gabbett (MCG) e de um método modificado de coloração a frio (MMC), ambos em duas etapas, em comparação com a do método de coloração fluorescente (MCF), em três etapas, para a detecção de BAAR em esfregaços de escarro. MÉTODOS: Nossa amostra consistiu de 260 amostras de escarro coletadas de casos suspeitos de tuberculose pulmonar no Kasturba Hospital, em Manipal, Índia. Os esfregaços foram preparados em triplicata, para cada um dos métodos: MCF, MMC e MCG. As lâminas foram numeradas aleatoriamente a fim de que o examinador fosse cegado quanto à identidade das amostras. RESULTADOS: Das 260 amostras, 16 (6,15 por cento), 15 (5,77 por cento) e 13 (5,00 por cento) foram positivas para BAAR com MCF, MMC e MCG, respectivamente. A sensibilidade de MCG e MMC em relação à de MCF foi de 81,25 por cento e 93,75 por cento, respectivamente. Houve boa concordância de MCG e MMC com MCF (0,988 e 0,996, respectivamente), e não houve diferenças estatísticas significativas. CONCLUSÕES: Embora MCG e MMC apresentaram menor sensibilidade que MCF, que é avaliado por microscopia de fluorescência, consideramos que os dois primeiros métodos sejam promissores no diagnóstico de tuberculose.

OBJECTIVE: In developing countries, sputum smear microscopy is the main tool for pulmonary tuberculosis case finding. The objective of the present study was to evaluate the diagnostic efficacy of Gabbett's staining (GS) and modified cold staining (MCS), both of which are two-step methods, in comparison with that of fluorescent staining (FS), which is a three-step method, for the detection of AFB in sputum smears. METHODS: Our sample comprised 260 sputum samples collected from individuals suspected of having pulmonary tuberculosis at Kasturba Hospital, in Manipal, India. Smears were prepared in triplicate: one each for FS, MCS, and GS. The smears were randomly numbered so that the examiner was blinded to the sample identities. RESULTS: Of the 260 samples, 16 (6.15 percent), 15 (5.77 percent), and 13 (5.00 percent) showed positive AFB results with FS, MCS, and GS, respectively. The sensitivity of GS and MCS, in comparison with that of FS, was 81.25 percent and 93.75 percent, respectively. The concordance of GS and MCS with FS was good (0.988 and 0.996, respectively), and no statistically significant differences were found. CONCLUSIONS: Although MCS and GS were found to be less sensitive than was FS, which is evaluated under fluorescence microscopy, the first two are promising methods for the diagnosis of tuberculosis.

Colletotrichum graminicola keratitis: First case report from India.

Colletotrichum graminicola is a medically important fungus belonging to the order Melanconiales under the class Coelomycetes. The members of the genus Colletotrichum are primarily plant pathogens which cause anthracnoses (fungal infection in plants). In the past few decades, they are progressively being implicated as etiological agents of subcutaneous hyalohyphomycoses and keratomycoses. Of the five medically important members in the genus Colletotrichum, keratitis due to Colletotrichum graminicola is rare. We diagnosed Colletotrichum graminicola keratitis in a 44-year-old man who presented with a non-healing corneal ulcer since three weeks. Positive smears and cultures from the corneal scrapings established the causative organism as C. graminicola. The patient was treated with a combination of oral ketoconazole and topical fluconazole and natamycin. Infection resolved over 10 weeks and antimicrobials were stopped. We describe the clinical presentation and treatment outcome of Colletotrichum graminicola keratitis.

Role of laboratory in rapid diagnosis of atypical mumps

Fairly large number of mumps virus infections present atypically without parotitis leading to delay in diagnosis and increased morbidity. Awareness of such presentations and inclusion of serological test for detecting IgM-specific antibodies could help in solving diagnostic dilemma, especially in unvaccinated individuals from developing countries.

Adequacy of two versus three sputum specimens for the diagnosis of pulmonary tuberculosis.

Background. The new guidelines issued by the Revised National Tuberculosis Control Programme for diagnosis of smear-positive tuberculosis recommend examination of only 2 sputum smears. We did a retrospective analysis of data from a designated microscopy centre to ascertain the diagnostic yield of 2 smears and the additional yield provided by the third smear. Methods. Data were obtained from the designated microscopy centre attached to our medical college. A total of 3257 patients with suspected tuberculosis had undergone sputum examination between September 2004 and March 2009. However, only 1762 of them had 3 sputum specimens examined. Data were entered and analysed using SPSS version 11.5. Results. Among the 1762 suspected patients, positivity in any 2 samples was found to be 17.7% while 19% were found to be positive in a single smear. A statistically insignificant association was found between the grading and positivity of the sputum samples using McNemar test. A positive third sample was found in 309 patients. If the first 2 samples were negative, the possibility of missing a third positive sample was 0.4%. Conclusion. Under field conditions, 2 sputum smears are as effective as 3 smears for diagnosing smear-positive tuberculosis.

Pulmonary nocardiosis due to nocardia farcinica in a renal transplant recipient.

Nocardia farcinica is an infrequent cause of nocardiosis among the renal transplant recipients and it has not been reported so far from India. We report a case of pulmonary nocardiosis due to N. farcinica in a 32-year-old woman with hypothyroidism and post-renal transplant status, currently on immunosuppressive therapy (prednisolone, azathioprine and tacrolimus). The N. farcinica isolate was susceptible to trimethoprim-sulfamethoxazole (TMP-SMZ), linezolid, imipenem, gentamicin but resistant to ceftriaxone, ciprofloxacin, tobramycin, erythromycin, amoxycillin-clavulanic acid and tetracycline. Treatment with TMP-SMZ and linezolid resulted in marked clinico-radiological improvement but after two weeks both of the drugs had to be stopped due to severe pancytopenia as adverse effect of their use. Currently, the patient is on imipenem and remains stable after four weeks of treatment. In N. farcinica infections, multi antibiotic resistance and toxicity of some specific drugs enhances the risk of therapeutic failure in renal transplant recipients.

Atypical presentations of melioidosis as emerging threat: a case report.

We report two atypical presentations of melioidosis as mediastinal lymphadenitis and prostatic abscess with Burkholderia pseudomallei, the emerging category 2 organism which led to diagnostic and therapeutic dilemma and thereby, delay in appropriate management. Any similar presentation should always be supported by microbiological opinion without any delay, which can help in instituting proper antibiotics with successful outcome.

Effect of "HAART" on CD4+ T lymphocyte counts in HIV seropositive south-Indian individuals--a follow up study.

Because of the drastic cut down in the prices of antiretroviral drugs, the antiretroviral therapy has become more affordable in India. 16 HIV-1 infected individuals who were on HAART were followed up for 12 months. The CD4 + T Cell count estimation was done at mean intervals of 6 months and 12 months. Antiretroviral therapy showed good response in 14(87.5%) and deterioration in 2 (12.5%) cases.

Evaluation of antibacterial activity of Mangifera indica on anaerobic dental microglora based on in vivo studies.

The present study was carried out to compare the subgingival plaque microflora in mango leaf users. Fifty subjects of both sexes, 25 of them used tooth brush and 25 used mango leaf as their home care hygiene device were included in the study. The microbiological evaluation for specific bacterial counts of Actinobacillus actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Fusobacterium nucleatum and Peptostreptococcus micros were carried out for all subject. Specific microbial evaluation revealed significant decrease in the proportion of P. intermedia and P. gingivalis in mango leaf users compared to tooth brush users. It shows that mangiferin possesses antibacterial activity in vivo against specific periodontal pathogens such as P. intermedia and P. gingivalis. Use of mango leaf in conjunction with a tooth brush will be a good home care device for maintenance of oral hygiene.