Dried blood spot HIV-1 RNA quantification: A useful tool for viral load monitoring among HIV-infected individuals in India.

Background & objectives: Monitoring of HIV-infected individuals on antiretroviral treatment (ART) ideally requires periodic viral load measurements to ascertain adequate response to treatment. While plasma viral load monitoring is widely available in high-income settings, it is rarely used in resource-limited regions because of high cost and need for sophisticated sample transport. Dried blood spot (DBS) as source specimens for viral load measurement has shown promise as an alternative to plasma specimens and is likely to be a useful tool for Indian settings. The present study was undertaken to investigate the performance of DBS in HIV-1 RNA quantification against the standard plasma viral load assay. Methods: Between April-June 2011, 130 samples were collected from HIV-1-infected (n=125) and non-infected (n=5) individuals in two district clinics in southern India. HIV-1 RNA quantification was performed from DBS and plasma using Abbott m2000rt system after manual RNA extraction. Statistical analysis included correlation, regression and Bland-Altman analysis. Results: The sensitivity of DBS viral load was 97 per cent with viral loads >3.0 log10 copies/ml. Measurable viral load (>3.0 log10 copies/ml) results obtained for the 74 paired plasma-DBS samples showed positive correlation between both the assays (r=0.96). For clinically acceptable viral load threshold values of >5,000 copies/ml, Bland-Altman plots showed acceptable limits of agreement (-0.21 to +0.8 log10 copies/ml). The mean difference was 0.29 log10 copies/ml. The cost of DBS was $2.67 lower compared to conventional plasma viral load measurement in the setting. Interpretation & conclusions: The significant positive correlation with standard plasma-based assay and lower cost of DBS viral load monitoring suggest that DBS sampling can be a feasible and economical means of viral load monitoring in HIV-infected individual in India and in other resource-limited settings globally.

In vitro production of biofilm in a flow cell system in a strain of Pseudomonas aeruginosa and Staphylococcus aureus and determination of efficiency of ciprofloxacin against them.

Background: Microorganisms develop biofilm on various medical devices. The process is particularly relevant in public health since biofilm associated organisms are much more resistant to antibiotics and have a potential to cause infections in patients with indwelling medical devices. Materials and Methods: To determine the efficiency of an antibiotic against the biofilm it is inappropriate to use traditional technique of determining Minimum Inhibitory Concentration (MIC) on the free floating laboratory phenotype. Thus we have induced formation of biofilm in two strains (Pseudomonas aeruginosa and Staphylococcus aureus, which showed heavy growth of biofilm in screening by Tube method) in a flow cell system and determined their antibiotic susceptibility against ciprofloxacin by agar dilution method in the range (0.25 mg/ml to 8 mg/ml). The MIC value of ciprofloxacin for the biofilm produced organism was compared with its free form and a standard strain as control on the same plates. Observations: Both the biofilm produced strains showed a higher resistance (MIC > 8 mg/ml) than its free form, which were 2 μg/ml for Pseudomonas aeruginosa and 4 mg/ml for Staphylococcus aureus. Thus biofilm can pose a threat in the patient treatment.

Community prevalence of methicillin and vancomycin resistant Staphylococcus aureus in and around Bangalore, southern India / Prevalência de Staphylococcus aureus resistente à meticilina e à vancomicina em comunidade no entorno de Bangalore, Índia do Sul

INTRODUCTION: Staphylococcus aureus is a known colonizer in humans and has been implicated in community acquired soft tissue infections. However emergence of methicillin resistant S. aureus (MRSA) has aroused great concern worldwide. This study aimed to determine the prevalence of MRSA in the community of Bangalore, southern India. METHODS: Swabs were collected from anterior nares, forearm, dorsum and palm of the hands of 1,000 healthy individuals residing in and around Bangalore, belonging to different socioeconomic strata and age groups. RESULTS: Analysis verified that 22.5 percent and 16.6 percent of the individuals presented Staphylococcus aureus and MRSA, respectively, at any of the three sites. Vancomycin resistance was observed in 1.4 percent of the S. aureus isolates, which was confirmed by detection of the vanA gene. It was interesting to note that 58.8 percent of the children in the age group 1-5 years-old presented MRSA, the highest percentage compared to other age groups of < 1 (44.4 percent) year-old, 5-20 (21.7 percent) years-old, > 40 (11 percent) years-old and 20-40 (9.9 percent) years-old. Among the population of various socioeconomic strata, maximum MRSA colonization was observed among doctors (22.2 percent), followed by upper economic class (18.8 percent), lower economic class (17.7 percent), apparently healthy hospital in-patients (16.5 percent), nurses (16 percent) and middle economic class (12.5 percent). Most of the MRSA isolates were capsular polysaccharide antigen type 8 (57.1 percent). CONCLUSIONS: There is a need for continuous surveillance and monitoring of the presence of MRSA in the community and a clearer understanding of the dynamics of the spread of MRSA will assist in controlling its dissemination.

INTRODUÇÃO: O Staphylococcus aureus é conhecido por ser um colonizador em humanos sendo implicado em infecções comunitárias dos tecidos moles. Contudo, a resistência à meticilina e emergência de S. aureus meticilina resistentes (MRSA) têm despertado preocupação em todo o mundo. O presente estudo visa encontrar a prevalência de MRSA na comunidade de Bangalore, sul da Índia. MÉTODOS: Suabes foram coletados de narinas anteriores, antebraço e dorso da palma de 1.000 indivíduos saudáveis, residentes em Bangalore e nas proximidades, pertencentes a diferentes estratos socioeconômicos e faixas etárias. RESULTADOS: Observou-se que 22,5 por cento e 16,6 por cento dos indivíduos foram abrigar Staphylococcus aureus e MRSA, respectivamente, em qualquer um dos três locais. Dos S. aureus isolados, 1,4 por cento também foram resistentes à vancomicina, o que foi confirmado pela detecção do gene vanA. Foi interessante notar que 58,8 por cento das crianças na faixa etária de 1-5 anos foram abrigar MRSA, o mais elevado em comparação com outros grupos etários de < 1 (44,4 por cento) ano, 50-20 (21,7 por cento) anos, > 40 (11 por cento) anos e 20-40 (9,9 por cento) anos. Entre a população de diferentes estratos socioeconômicos, a colonização de MRSA máxima foi observada entre os médicos (22,2 por cento), seguida pela classe econômica superior (18,8 por cento), classe baixa (17,7 por cento), pacientes aparentemente saudáveis (16,5 por cento), enfermeiros (16 por cento) e classe econômica média (12,5 por cento). A maioria dos MRSA isolados eram do tipo polissacarídeo capsular antígeno 8 (57,1 por cento). CONCLUSÕES: Há uma necessidade de vigilância e monitorização contínua da presença de MRSA na comunidade, bem como uma melhor compreensão da dinâmica de propagação de MRSA pode ajudar no controle da disseminação.

Ceftriaxone resistant Shigella Flexneri, an emerging problem.

Shigellosis is a disease of public health importance in developing countries. It may cause self-limited diarrhea to severe dysentery. Emergence of multi drug resistant (MDR) strains is a growing concern globally. Ceftriaxone and ciprofloxacin are the drugs of choice for MDR cases. Here, we report a case of MDR Shigella flexneri from an immunocompromised patient. The strain was resistant to ceftriaxone [minimum inhibitory concentration (MIC) ≥ 64 μg/ml], limiting the treatment option. Simultaneously, the strain was also found to be resistant to ciprofloxacin (MIC ≥ 4 μg/ml). However, it was susceptible to ceftazidime (MIC 4 μg/ml). This is the first case of ceftriaxone resistant Shigella spp. reported from our hospital.

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.