A toolbox for tuberculosis diagnosis: an Indian multicentric study (2006-2008): microbiological results

Background

The aim of this multicentric prospective study in India was to assess the value of several microbiological tools that contribute to the diagnosis of tuberculosis (TB) according to HIV status.

Methods

Standard microbiological tools on individual specimens were analyzed.

Results

Among the 807 patients with active TB, 131 were HIV-infected, 316 HIV-uninfected and 360 had HIV-unknown status. Among the 980 non-active TB subjects, 559 were at low risk and 421 were at high risk of M. tuberculosis (Mtb) exposure. Sensitivity of smear microscopy (SM) was significantly lower in HIV-infected (42.2%) than HIV-uninfected (75.9%) (p = 0.0001) and HIV-unknown pulmonary TB patients (61.4%) (p = 0.004). Specificity was 94.5% in non-TB patients and 100% in health care workers (HCW) and healthy family contacts. Automated liquid culture has significantly higher diagnostic performances than solid culture, measured by sensitivity (74.7% vs. 55.9%) (p = 0.0001) and shorter median time to detection (TTD) (12.0 vs. 34.0 days) (p = 0.0001). Specificity was 100% in HCW and cured-TB patients, but was lower in non-TB patients (89%) due to isolation of Mycobacteria other than tuberculosis (MOTT). TTD by both methods was related to AFB score. Contamination rate was low (1.4%). AccuProbe hybridization technique detected Mtb in almost all culture-positive specimens, but MOTT were found in 4.7% with a significantly higher frequency in HIV-infected (15%) than HIV-uninfected TB patients (0.5%) (p = 0.0007). Pre-test classification significantly increased the diagnostic value of all microbiological tests in pulmonary TB patients (p<0.0001) but to a lesser degree in extrapulmonary TB patients.

Conclusions

Conventional microbiological tools led to results similar to those already described in India special features for HIV-infected TB patients included lower detection by SM and culture. New microbiological assays, such as the automated liquid culture system, showed increased accuracy and speed of detection.     

Cytologic diagnosis of leishmaniasis in HIV infection. A report of eight cases

BACKGROUND: Leishmania organisms are among the intracellular microorganisms with a tendency to develop in patients with the acquired immunodeficiency syndrome (AIDS). With increasing travel to endemic areas by patients with human immunodeficiency virus (HIV) infection, it is becoming a more-frequent diagnosis in nonendemic areas. CASES: Ten cytologic specimens from eight patients with leishmaniasis and AIDS were reviewed. Eight samples were obtained from lymph nodes through fine needle aspiration (FNA). Another sample was obtained after scraping a tongue ulcer. The last one was an ascitic fluid specimen. Smears showed numerous parasitized histiocytes with abundant intracellular Leishmania organisms (amastigotes). Extracellular microorganisms were also abundant. Diff-Quik-stained smears allowed the clear recognition of the characteristic morphologic appearance with a deep-staining area (nuclei) and paranuclear zone (kinetoplast). Intracellular organisms were round, while single, extracellular forms were a more elongated. CONCLUSION: The polymorphous clinical manifestations usually seen in patients suffering from leishmaniasis and AIDS constitute a diagnostic challenge that can be facilitated by cytopathologic examination. Cytology permits easy and rapid identification of Leishmania amastigotes, allowing a specific diagnosis and treatment.     

Improving the quality of care via maintenance of certification and the Web: an early status report

Few question the need for continuous professional development throughout a physician's career, but rapid changes in health care are creating demand for physicians to acquire new knowledge, skills, and attitudes to implement quality improvement in clinical practice. The Internet and World Wide Web are technologies that have the potential to facilitate deep change in physician practice and lifelong learning. This paper describes how the American Board of Internal Medicine (ABIM) has utilized the Web and the Internet to engage physicians in the competencies of practice-based learning and improvement and systems-based practice. Specifically, we describe how the ABIM developed and implemented Web-based practice improvement modules (PIMs) to help physicians measure and improve their clinical practice.     

Morbidity and risk of subsequent diagnosis of HIV: a population based case control study identifying indicator diseases for HIV infection

Background

Early identification of persons with undiagnosed HIV infection is an important health care issue. We examined associations between diseases diagnosed in hospitals and risk of subsequent HIV diagnosis.

Methods

In this population-based case control study, cases were persons with incident HIV infection diagnosed in Denmark between 1 January 1995 and 1 June 2008. Risk-set sampling was used to identify 19 age- and gender-matched population controls for each HIV case, using the HIV diagnosis date as the index date for both cases and controls. Prior hospital diagnoses obtained from Danish medical databases were first categorized into 22 major disease categories (excluding AIDS-defining diseases except tuberculosis) and then subdivided into 161 subcategories, allowing us to examine specific diseases as potential HIV indicators by conditional logistic regression.

Results

The study included 2,036 HIV cases and 35,718 controls. Persons with the following disease categories had a high risk of HIV diagnosis during the subsequent 5-year period: sexually transmitted infections and viral hepatitis (adjusted odds ratio [aOR] = 12.3, 95% CI: 9.60–15.7), hematological diseases (aOR = 4.28, 3.13–5.85), lower respiratory tract infections (aOR = 3.98, 3.14–5.04)), CNS infections (aOR = 3.44, 1.74–6.80), skin infections (aOR = 3.05, 2.47–3.75), other infections (aOR = 4.64, 3.89–5.54), and substance abuse (aOR = 2.60, 2.06–3.29). Several specific diseases were associated with aORs >20 including syphilis, hepatitis A, non “A” viral hepatitis, herpes zoster, candida infection, endocarditis, thrombocytopenia, and opioid abuse.

Conclusions

Targeted testing for HIV in patients diagnosed with diseases associated with HIV may lead to earlier treatment and thereby reduced morbidity, mortality and HIV transmission.     

First Italian case of cyclosporiasis in an immunocompetent woman: local acquired infection

Cyclospora cayetanensis is a coccidian agent of chronic diarrhea in humans with a worldwide distribution. We report the first documented case of acquired Cyclosporiasis in Italy. The patient was an immunocompetent woman with no recent history of travel outside the country. Microscopy detected Cyclospora oocysts in a feces sample. PCR detected the pathogen in a second sample, which had tested negative by microscopy. The patient was investigated to detect other microorganisms in feces, such as Salmonella spp., Shigella spp, Campylobacter spp., Yersinia spp, and enteroviruses: all were negative. All symptoms disappeared 72 h after the beginning of therapy.     

The principles of pre-epidemic diagnosis in the epidemiological surveillance system for HIV infection

The problem of the general introduction of the principles of the pre-epidemic diagnostics of HIV infection was discussed. These principles should be based on the observation of "model" infections (viral hepatitides B, C, D), multipurpose serological monitoring and sociological methods aimed at obtaining the necessary information. The suggestion that the age and sexual structure of HIV-infected patients would be determined by the route of HIV transmission, prevailing on a given territory, including into infection process adolescents and young women and men was made.     

The 2001-03 Famine and the Dynamics of HIV in Malawi: A Natural Experiment

Background

Food security has deteriorated for many people in developing regions facing high and volatile food prices. Without effective and equitable responses, the situation is likely to worsen due to diminishing access to land and water, competition from non-food uses of agricultural products, and the effects of climate change and variability. Understanding how this will affect the burden and distribution of major diseases such as HIV is critical. This study makes use of the near-experimental conditions created by the Malawi famine to shed new light on this issue.

Methods

Multilevel, random intercept models were used to relate the change in HIV prevalence at antenatal surveillance sites over the course of the famine to the proportion of rural households requiring food aid in the surrounding district at the famine’s peak. Similar models were used to relate this indicator of rural hunger to changes in the composition of the antenatal population. The extent and direction of migration were estimated from a household survey conducted 1–2 years after the famine.

Findings

At rural sites, the change in HIV prevalence was positively and non-linearly related to the extent of rural hunger (P = 0.016), consistent with contemporary accounts of increased transactional sex and with hunger compromising immune function. At non-rural sites, prevalence declined as rural hunger increased (P = 0.006), concentrated in women who self-identified as farmers (P = 0.010). This finding is consistent with contemporary accounts of migration in search of food and work from villages where HIV risk was lower to towns and cities where it was higher. Corroborating this interpretation, the proportion of farmers in the antenatal population was found to rise at non-rural sites as rural hunger increased in the surrounding district (P = 0.015) whereas the proportion fell with increasing rural hunger at rural sites (P<0.001). The models suggest migrants were predominantly farming women under 25 years (P = 0.010). The household survey confirmed that there was a surge of rural-to-urban migration during the famine, particularly by women under 25 years. Migration to less affected rural areas also increased.

Conclusion

The Malawi famine appears to have had a substantial effect on HIV’s dynamics and demography. Poverty and inequality, commonly considered structural determinants of HIV epidemics, can change rapidly, apparently transmitting their effects with little lag. Epidemic patterns risk being misread if such social and economic change is ignored. Many studies examining HIV prevalence declines have implicated sexual behaviour change but do not appear to have adequately considered the contribution of rural-urban migration. The evidence from Malawi, which links actions that undermined people’s food security to changes in the prevalence and distribution of HIV infections, suggests new opportunities for prevention.     

Production of IgM specific recombinant dengue multiepitope protein for early diagnosis of dengue infection

Dengue virus infections have recently undergone dramatic expansion in range, affecting several tropical and subtropical regions of the world. Early detection of dengue infection based on the identification of antibodies has emerged as a practical and reliable means of diagnosis of dengue fever. The recombinant dengue multiepitope (rDME-M) protein specific to IgM in E. coli was produced in a 5-L fermentor for use in diagnostic purpose. After fermentation, dry cell weight was approximately 11.8 g/L of the culture. The rDME-M protein was purified under denaturing conditions using single-step nickel nitrilotriacetate (Ni-NTA) affinity chromatography. The final yield of purified rDME-M protein from this method was approximately 68.5 mg/L of the culture. The purity of rDME-M protein was checked by SDS-PAGE analysis, and the reactivity of this protein was further checked by Western blotting and enzyme-linked immunosorbent assay (ELISA). The purified protein was used as an antigen in the development of an in-house dipstick ELISA and evaluated with a panel of 80 patient sera, characterized using commercially available tests for detection of dengue antibody. The results were in excellent agreement with those of IgM capture ELISA (Pan-Bio) and rapid immunochromatography (IC) test (Pan-Bio). These results show that the in-house dipstick ELISA using rDME-M protein can be used as a promising kit because of its comparable sensitivity, specificity, field applicability, and low cost.     

Using PCR for early diagnosis of bovine leukemia virus infection in some native cattle

Bovine leukemia virus (BLV), the causative agent of enzootic bovine leukosis, is an exogenous, B lymphotropic retrovirus belonging to the Retroviridae family that induces persistent lymphocytosis in cattle and sheep. PCR has proven to be particularly suitable for investigating herds of cattle with a very low incidence of BLV infection and for clarifying doubtful serological results obtained by immunodiffusion or ELISA. The native Iranian and Russian cattle have a series of valuable traits that discriminate them as unique breeds that are well able to compete with western analogues. However, their gene pools have not been analyzed with molecular markers, including detection of BLV by PCR. Two pairs of primers were used: gag1 and gag2, and pol1 and pol2, which encompass 347- and 599-bp fragments of the BLV gene, respectively. Sixty-five Iranian Sistani, 120 Yaroslavl, 50 Mongolian, and 35 Black Pied cows were investigated. Among these 270 animals, we obtained 42 positive and 15 doubtful results in the first PCR. The second PCR was very effective in increasing BLV test reliability data to support detection of BLV.     

Fine needle aspiration cytology determinants of the diagnosis of primary nodal Kaposi's sarcoma as the first sign of unknown HIV infection: a case report

BACKGROUND: Kaposi's sarcoma (KS) is a vascular malignant tumor characterized by human herpesvirus 8 infection of neoplastic cells. Diffuse cutaneous lesions represent the classical clinical presentation. This case report describes the first fine needle aspiration cytology findings of a primary lymph nodal KS, a rather unusual localization of the disease. CASE: A 28-year-old, apparently healthy man saw a surgeon for right inguinal node enlargement without other symptoms. The clinician performed fine needle aspiration and made a preliminary diagnosis of a neoplasm of probable mesenchymal origin, not otherwise specified. The lymph node was excised, and the final histologic diagnosis was primary lymphoadenopathic KS. A serologic test revealed antibody positivity for HIV. CONCLUSION: The diagnosis of primary KS of the lymph node, in the absence of any other clinical manifestation, was the first sign of HIV infection.