Human leptospirosis in the Caribbean, 1997-2005: characteristics and serotyping of clinical samples from 14 countries.

OBJECTIVE: To determine the frequency of human leptospirosis in the sera of suspected clinical cases sent by 14 Caribbean countries for diagnosis to a regional laboratory in 1997-2005. METHODS: All serum samples were initially tested using the immunoglobulin M (IgM) enzyme-linked immunosorbent assay (ELISA) for leptospirosis. Demographic data (such as age and sex), month of the year and clinical manifestations that had been observed by the attending physician were related to seropositivity. The microscopic agglutination test (MAT) was used to serotype sera using a panel of 23 international serovars. RESULTS: Of 3 455 samples tested, 452 (13.1%) were seropositive for IgM antibodies to leptospirosis by the ELISA, with frequencies significantly (P < 0.05; χ2) different across countries and years. Among seropositive patients, the frequency of detection of leptospirosis (23.1%) was significantly higher in the age groups 1-20 years and 31-40 years combined compared with other age groups; and in male patients (72.1%) compared with female patients (19.7%) (P < 0.05; χ2). Chills, jaundice, vomiting, weakness, diarrhea, and kidney failure/problems were significantly (P < 0.05; χ2) exhibited at a higher frequency in seropositive, rather than seronegative patients. Using the MAT on 100 sera tested, 98 (98%) were seropositive, of which the serogroup Icterohaemorrhagiae was most prevalent with the detection of serovars Copenhageni (70%), Icterohaemorrhagiae (67%), and Mankarso (29%). CONCLUSIONS: Since only 13.1% of the suspected cases of leptospirosis were seropositive for IgM ELISA antibodies, other clinical conditions may have been responsible for the clinical manifestations observed, or the patient may have had chronic leptospirosis (IgG). In the Caribbean, serovars of the serogroup Icterohaemorrhagiae were responsible for most infections in the cases tested.

Comparison of the QuantiFERON®-TB Gold assay and tuberculin skin test to detect latent tuberculosis infection among target groups in Trinidad & Tobago.

OBJECTIVE: To compare the QuantiFERON®-TB Gold (QFT-G) assay and tuberculin skin test (TST) in screening/diagnosis of latent tuberculosis infection (LTBI) among individuals in Trinidad & Tobago at high risk for TB. METHODS: A total of 560 individuals (TB patient contacts, HIV patients, health care workers, prison inmates, and TB patients [controls]) were recruited for the study. Blood was drawn and processed using the QFT-G assay, followed by immediate administration of TST solution on subjects' forearm. Data were analyzed with Epi InfoTM 3.5.1 software. Results were compared across the target groups using the chi-square test (P < 0.05). RESULTS: The QFT-G assay detected LTBI in 51% of the subjects (with most positive results occurring among the control group) whereas the TST detected it in 39.4% (P = 0.001). Overall, the QFT-G assay detected LTBI more frequently than the TST among all subjects except the control group, where detection favored the TST. The QFT-G assay produced indeterminate and nonreactive results in some HIV patients but required less turnaround time than the TST (23.3 h versus 70.2 h; P < 0.0001). The TST cost less per subject than the QFT-G assay (US $3.70 versus US $18.60; P = 0.0008). CONCLUSIONS: The QFT-G assay cost more but had a higher detection rate among most target groups and required less turnaround time than the TST. However, its sensitivity was lower among immunocompromised subjects. Therefore, the QFT-G assay should be used with caution for LBTI screening/diagnosis in resource-poor, high-HIV prevalence settings such as Trinidad & Tobago.

First insight into Mycobacterium tuberculosis epidemiology and genetic diversity in Trinidad and Tobago.

This report is based on a 1-year recruitment of all of the culture-positive Mycobacterium tuberculosis cases in Trinidad and Tobago (n = 132). The study population was characterized by a high male-to-female sex ratio of 4 and a human immunodeficiency virus-tuberculosis (TB) coinfection rate of 30%. It mainly occurred among African descendants, who represent 37.5% of the total population but 69.7% of all TB cases (P < 0.001). Spoligotyping resulted in 25 different patterns and 12 clusters (2 to 74 strains per cluster), with the predominance of a highly conserved spoligotype international type clone, SIT566.

Evaluation of methods for rapid detection of resistance to isoniazid and rifampin in Mycobacterium tuberculosis isolates collected in the Caribbean.

The rapid identification of drug-resistant strains of Mycobacterium tuberculosis is crucial for the timely initiation of appropriate antituberculosis therapy. The performance of the Genotype MTBDRplus assay was compared with that of the Bactec 460 TB system, a "gold standard" culture-based method. The Genotype MTBDRplus assay was quicker and more cost-effective for the detection of rifampin resistance, but it was not as good for the detection of isoniazid-resistant strains in our setting.

Finer characterization of Mycobacterium tuberculosis using spoligotyping and 15-loci MIRU-VNTRs reveals phylogeographical specificities of isolates circulating in Guyana and Suriname.

In this study we used spoligotyping and 15-loci MIRU-VNTRs for a finer characterization of Mycobacterium tuberculosis strains isolated from patients residing in Guyana (n=74) and Suriname (n=80). The mean age of the patients was 38.5years (36.5 and 40.2years for Guyana vs. Suriname), with a male-to-female sex-ratio of 2.25 for Guyana vs. 4.27 in Suriname. Spoligotyping and 15-loci MIRU-VNTRs led to a total of 41 and 65 different patterns respectively, with an overall clustering rate of 83.8% vs. 68.8%. Combined spoligotyping and VNTR analysis led to the detection of 18 clusters of 2-41 isolates, with an overall clustering of 67.5% and a recent "n-1" transmission rate of 55.8%. Importantly, Guyana was characterized by a significantly higher percentage of clustered isolates than Suriname (79.7% vs. 56.3%; p=0.0019). Three big spoligo/MIRU (SIT/MIT) clusters containing >10 isolates each were shared between the 2 countries, and concerned: T1 sublineage cluster 53/861 (n=41, 37 in Guyana vs. 4 in Suriname); EAI6-BGD1 sublineage cluster 1340/860 (n=16, 3 in Guyana vs. 13 in Suriname); and T1 sublineage cluster 131/146 (n=11, 6 in Guyana vs. 5 in Suriname); as well as 2 smaller clusters of 2 and 3 isolates respectively. However, the relative phylogeographical specificities of strains in circulation as well as a lack of drug-resistance observed among strains from Suriname suggested that trans-border transmission of drug-resistant isolates occurred less frequently than thought. Tracing and interrupting transmission channels of a specific clone (SIT53/15-MIT861) should become a priority in Guyana, not only because it is by far most abundant but also because it accounts for almost half of the drug resistant isolates (n=8/17, 47.1%) in our study, and clustered 5/12 (41.7%) MDR isolates.

Seroprevalence of toxocariasis in schoolchildren in Trinidad.

The seroprevalence of toxocariasis was investigated in 1997/98 in 1009 schoolchildren (aged 5-12 years) throughout Trinidad. Infection, as measured by titre, was found to be high compared to values obtained from children in other countries. Using an excretory-secretory antigen and performing an ELISA test, it was found that 62.3% of children had an IgG antibody titre of > or = 1:100, indicating exposure to the parasite, while 27.2% had a titre of > or = 1:800, indicating a current or recent infection. Relationships were explored between seroprevalence and host factors including age, sex, school location, and other risk factors including geophagia, thumb-sucking, presence of other gastrointestinal-tract parasitism and pet ownership. There was no significant relationship between age and the presence of current or recent infection (P = 0.746). Boys were significantly more commonly infected than girls as were the attendees of rural schools versus urban schools (P < 0.001). The percentage of seropositivity among children varied widely from school to school. Pet ownership and the absence of pipe-borne water in the household were found to be significantly associated with positive serology (P < 0.05). Clinical symptoms mostly associated with positive serology were eczema, seizures and chronic cough. Recommendations derived from this study include health education in order to increase the public awareness on the transmission of the disease, de-worming all dogs and cats periodically and the curbing of stray dogs and cats. Environmental sanitation measures should include keeping children away from contaminated areas and practising proper hygiene after play.

Evidence for the interruption of transmission of lymphatic filariasis among schoolchildren in Trinidad and Tobago.

This study was carried out to provide some evidence for the interruption of transmission of lymphatic filariasis (LF) among schoolchildren in Trinidad and Tobago. A cross-sectional survey for LF antigenaemia was performed among 63 (13.2%) of the 479 primary schools located in eight administrative (and geographical) regions of Trinidad and Tobago. From these communities, 2597 schoolchildren aged 6-12 years were sequentially selected for a survey of bancroftian antigenaemia. From each child, 100 microl (finger-prick) whole blood sample was applied to a Binax immunochromatographic card test (ICT), and read for the presence of antigenaemia. The ICT results showed a negative finding for LF antigenaemia and suggest that LF transmission has been interrupted in the survey areas.

A first assessment of Mycobacterium tuberculosis genetic diversity and drug-resistance patterns in twelve Caribbean territories.

With the exception of some French-speaking islands, data on tuberculosis (TB) in the Caribbean are scarce. In this study, we report a first assessment of genetic diversity of a convenience sample of Mycobacterium tuberculosis strains received from twelve Caribbean territories by spoligotyping and describe their drug-resistance patterns. Of the 480 isolates, 40 (8.3%) isolates showed resistance to at least one anti-TB drug. The proportion of drug-resistant strains was significantly higher in The Bahamas (21.4%; P = 0.02), and Guyana (27.5%; P < 0.0001), while it was significantly lower in Jamaica (2.4%; P = 0.03) than in other countries of the present study. Regarding genetic diversity, 104 distinct spoligotype patterns were observed: 49 corresponded to clustered strains (2 to 93 strains per cluster), while 55 remained unclustered among which 16 patterns were not reported previously. Combining the study results with regional data retrieved from the international SITVIT2 database underlined a connection between frequency of certain M. tuberculosis phylogenetic lineages and the language spoken, suggesting historical (colonial) and ongoing links (trade, tourism, and migratory flows) with European countries with which they shared a common past.

Improving quality in national reference laboratories: The role of SLMTA and mentorship.

BACKGROUND: The Nigerian Institute of Medical Research houses two reference laboratories: the virology and tuberculosis laboratories. Both were enrolled in the Strengthening Laboratory Management Toward Accreditation (SLMTA) programme. OBJECTIVE: To describe the impact of SLMTA and discuss factors affecting the results, with an emphasis on mentorship. METHODS: The SLMTA programme was implemented from April 2010 through November 2012. Participants attended three workshops and executed quality improvement projects; laboratory auditors evaluated performance using a standard checklist. The virology laboratory did not receive mentorship; however, the tuberculosis laboratory had an international mentor who visited the laboratory four times during the programme, spending two to four weeks embedded within the laboratory during each visit. RESULTS: There was an overall improvement in the performance of both laboratories, with the virology laboratory increasing 13% (from 80% at baseline to 93% at exit audit) and the tuberculosis laboratory increasing 29% (from 66% to 95%). These scores were maintained nine months later at the surveillance audit. CONCLUSION: The SLMTA programme resulted in improved and sustained quality management performance for both laboratories. Mentoring was a possible factor in the substantial improvement made by the tuberculosis laboratory and should be considered in order to augment the training received from the SLMTA workshops.

Leptospirosis outbreak following severe flooding: a rapid assessment and mass prophylaxis campaign; Guyana, January-February 2005.

BACKGROUND: Leptospirosis is a zoonosis usually transmitted through contact with water or soil contaminated with urine from infected animals. Severe flooding can put individuals at greater risk for contracting leptospirosis in endemic areas. Rapid testing for the disease and large-scale interventions are necessary to identify and control infection. We describe a leptospirosis outbreak following severe flooding and a mass chemoprophylaxis campaign in Guyana. METHODOLOGY/PRINCIPAL FINDINGS: From January-March 2005, we collected data on suspected leptospirosis hospitalizations and deaths. Laboratory testing included anti-leptospiral dot enzyme immunoassay (DST), immunohistochemistry (IHC) staining, and microscopic agglutination testing (MAT). DST testing was conducted for 105 (44%) of 236 patients; 52 (50%) tested positive. Four (57%) paired serum samples tested by MAT were confirmed leptospirosis. Of 34 total deaths attributed to leptospirosis, postmortem samples from 10 (83%) of 12 patients were positive by IHC. Of 201 patients interviewed, 89% reported direct contact with flood waters. A 3-week doxycycline chemoprophylaxis campaign reached over 280,000 people. CONCLUSIONS: A confirmed leptospirosis outbreak in Guyana occurred after severe flooding, resulting in a massive chemoprophylaxis campaign to try to limit morbidity and mortality.

Phylogeographical and molecular characterization of an emerging Mycobacterium tuberculosis clone in Trinidad and Tobago.

We report on a fine molecular and phylogenetical characterization of circulating Mycobacterium tuberculosis strains isolated from patients during a 1-year period in Trinidad and Tobago (T&T). The spoligotyping data coupled to minisatellite typing and available epidemiological data showed that a single major clone of "evolutionary modern" tubercle bacilli (SIT566) was responsible for more than half of the tuberculosis (TB) cases. It preferentially infected younger age groups (mean 39.1 years versus 47.7 years for other genotypes, p<0.0005), and was overrepresented in Port-of-Spain (1 out of 3 patients). A comparison of genotyping results to data gathered for 6 Caribbean countries (n=2653 clinical isolates) showed that the overall lineage distribution in T&T was completely different from its neighbors, e.g., T&T was the only country harboring a unique sublineage of the Latin American & Mediterranean (LAM) family, designated LAM-10CAM with phylogeographical specificity for Cameroon and neighboring countries in West Africa; interestingly 3/4 of the patients within this group in T&T were African descendants. Similarly, strains belonging to East African Indian (EAI) lineage with phylogeographical specificity for the Indian subcontinent, were found in T&T (13% of all strains), but were absent among the neighboring countries. Although the predominant SIT566 was not yet detected elsewhere in the Caribbean, available information underlined that this genotype was already present in the United States as imported cases of disease among T&T-born patients. Characterization of SIT566 strains using 12-, 15- and 24-loci MIRU typing, and comparison of results to international databases showed that these isolates were characterized by a common 12-loci MIRU pattern 224315153324 corresponding to MIRU International Type-MIT633 in 21/25 strains tested, as well as its 4 variants; an orphan pattern , MIT27-, MIT117-, and MIT1158-. Extended 24-loci MIRU typing led to a predominant pattern 224315153324323483334323 in a total of 16/21 MIT633 isolates, as well as identification of 3 supplemental patterns. Comparison of 24-loci MIRU data with the international database MIRU-VNTRplus showed the unique nature of the patterns obtained in T&T. Further analysis using the Levenshtein algorithm showed that the first 2 closest matches with the SIT566/MIT633 clone belonged to the X lineage strains in MIRU-VNTRplus. This observation corroborates our preliminary spoligotyping-based analysis using minimum spanning and neighbor-joining trees, which suggested a phylogenetical relatedness of the SIT566 clone with SIT119, which represents X1 lineage prototype in SpolDB4 database. We hypothesize that the predominant SIT566 clone might have evolved from a pool of X lineage M. tuberculosis strains with phylogeographical affinity for Anglo-Saxon descendants.

Human leptospirosis in the Caribbean, 1997-2005: characteristics and serotyping of clinical samples from 14 countries / Leptospirosis humana en el Caribe entre 1997 y 2005: características y serotipificación de muestras clínicas de 14 países

OBJECTIVE: To determine the frequency of human leptospirosis in the sera of suspected clinical cases sent by 14 Caribbean countries for diagnosis to a regional laboratory in 1997-2005. METHODS: All serum samples were initially tested using the immunoglobulin M (IgM) enzyme-linked immunosorbent assay (ELISA) for leptospirosis. Demographic data (such as age and sex), month of the year and clinical manifestations that had been observed by the attending physician were related to seropositivity. The microscopic agglutination test (MAT) was used to serotype sera using a panel of 23 international serovars. RESULTS: Of 3 455 samples tested, 452 (13.1 percent) were seropositive for IgM antibodies to leptospirosis by the ELISA, with frequencies significantly (P < 0.05; χ2) different across countries and years. Among seropositive patients, the frequency of detection of leptospirosis (23.1 percent) was significantly higher in the age groups 1-20 years and 31-40 years combined compared with other age groups; and in male patients (72.1 percent) compared with female patients (19.7 percent) (P < 0.05; χ2). Chills, jaundice, vomiting, weakness, diarrhea, and kidney failure/problems were significantly (P < 0.05; χ2) exhibited at a higher frequency in seropositive, rather than seronegative patients. Using the MAT on 100 sera tested, 98 (98 percent) were seropositive, of which the serogroup Icterohaemorrhagiae was most prevalent with the detection of serovars Copenhageni (70 percent), Icterohaemorrhagiae (67 percent), and Mankarso (29 percent). CONCLUSIONS: Since only 13.1 percent of the suspected cases of leptospirosis were seropositive for IgM ELISA antibodies, other clinical conditions may have been responsible for the clinical manifestations observed, or the patient may have had chronic leptospirosis (IgG). In the Caribbean, serovars of the serogroup Icterohaemorrhagiae were responsible for most infections in the cases tested.

OBJETIVO: Determinar la frecuencia de leptospirosis humana en el suero de presuntos casos clínicos enviados por 14 países del Caribe a un laboratorio regional para la confirmación del diagnóstico entre 1997 y 2005. MÉTODOS: Todas las muestras de suero se analizaron inicialmente mediante el ensayo inmunoenzimático de adsorción (ELISA) para detectar inmunoglobulina M (IgM) contra Leptospira. Se relacionó la seropositividad con datos demográficos (como la edad y el sexo), el mes del año y las manifestaciones clínicas observadas por el médico a cargo. Se usó la prueba de aglutinación microscópica para serotipificar los sueros con un grupo de 23 serovariedades internacionales. RESULTADOS: De las 3 455 muestras analizadas por ELISA, 452 (13,1 por ciento) fueron seropositivas para anticuerpos IgM contra Leptospira, con frecuencias significativamente diferentes (P < 0,05; χ2) según el país y el año. En los pacientes seropositivos, la frecuencia de detección de leptospirosis (23,1 por ciento) fue significativamente mayor en los grupos etarios de 1 a 20 años y de 31 a 40 años combinados, en comparación con otros grupos de edad; y mayor en los varones (72,1 por ciento) en comparación con las mujeres (19,7 por ciento) (P < 0,05; χ2). Los escalofríos, la ictericia, los vómitos, la debilidad, la diarrea y la insuficiencia o los trastornos renales fueron significativamente más frecuentes (P < 0,05; χ2) en los pacientes seropositivos que en los seronegativos. De los 100 sueros que se analizaron con la prueba de aglutinación microscópica, 98 (98 por ciento) fueron seropositivos, y entre estos el serogrupo Icterohaemorrhagiae fue el más frecuente, con detección de las serovariedades Copenhageni (70 por ciento), Icterohaemorrhagiae (67 por ciento) y Mankarso (29 por ciento). CONCLUSIONES: Ya que solo 13,1 por ciento de los presuntos casos de leptospirosis fueron seropositivos por ELISA para anticuerpos IgM, las manifestaciones clínicas observadas pueden haberse debido a otras enfermedades, o el paciente puede haber tenido leptospirosis crónica (con anticuerpos IgG). En los casos analizados en el Caribe, las serovariedades del serogrupo Icterohaemorrhagiae causaron la mayoría de las infecciones.

Comparison of the QuantiFERON®-TB Gold assay and tuberculin skin test to detect latent tuberculosis infection among target groups in Trinidad & Tobago / Comparación entre la prueba QuantiFERON®-TB Gold y la prueba cutánea de la tuberculina para detectar la infección tuberculosa latente en grupos destinatarios en Trinidad y Tabago

OBJECTIVE: To compare the QuantiFERON®-TB Gold (QFT-G) assay and tuberculin skin test (TST) in screening/diagnosis of latent tuberculosis infection (LTBI) among individuals in Trinidad & Tobago at high risk for TB. METHODS: A total of 560 individuals (TB patient contacts, HIV patients, health care workers, prison inmates, and TB patients [controls]) were recruited for the study. Blood was drawn and processed using the QFT-G assay, followed by immediate administration of TST solution on subjects' forearm. Data were analyzed with Epi InfoTM 3.5.1 software. Results were compared across the target groups using the chi-square test (P < 0.05). RESULTS: The QFT-G assay detected LTBI in 51 percent of the subjects (with most positive results occurring among the control group) whereas the TST detected it in 39.4 percent (P = 0.001). Overall, the QFT-G assay detected LTBI more frequently than the TST among all subjects except the control group, where detection favored the TST. The QFT-G assay produced indeterminate and nonreactive results in some HIV patients but required less turnaround time than the TST (23.3 h versus 70.2 h; P < 0.0001). The TST cost less per subject than the QFT-G assay (US $3.70 versus US $18.60; P = 0.0008). CONCLUSIONS: The QFT-G assay cost more but had a higher detection rate among most target groups and required less turnaround time than the TST. However, its sensitivity was lower among immunocompromised subjects. Therefore, the QFT-G assay should be used with caution for LBTI screening/diagnosis in resource-poor, high-HIV prevalence settings such as Trinidad & Tobago.

OBJETIVO: Comparar la prueba QuantiFERON®-TB Gold (QFT-G) con la prueba cutánea de la tuberculina (PPD) para el tamizaje y diagnóstico de la infección tuberculosa latente (ITBL) en personas con alto riesgo de tuberculosis en Trinidad y Tabago. MÉTODOS: Para el estudio, se reclutó un total de 560 individuos (personas en contacto con pacientes de tuberculosis, pacientes con VIH, trabajadores de la salud, presidiarios y pacientes de tuberculosis [grupo testigo]). Las muestras de sangre se extrajeron y procesaron utilizando la prueba QFT-G, seguida de la aplicación inmediata de la solución de PPD en el antebrazo de las personas. Los datos se analizaron con el software Epi InfoTM 3.5.1. Los resultados obtenidos en los grupos destinatarios se compararon utilizando la prueba de la ji al cuadrado (P < 0,05). RESULTADOS: La prueba QFT-G detectó la infección tuberculosa latente en 51 por ciento de los individuos (la mayoría de los resultados positivos se presentaron en el grupo testigo) mientras que la prueba PPD la detectó en 39,4 por ciento (P = 0,001). En términos generales, la prueba QFT-G detectó la infección tuberculosa latente con mayor frecuencia que la PPD en todos los individuos, excepto en aquellos del grupo testigo, donde el índice de detección favoreció a la PPD. La prueba QFT-G produjo resultados indeterminados y no reactivos en algunos pacientes con VIH, pero requirió menos tiempo de respuesta que la PPD (23,3 h contra 70,2 h; P < 0.0001). La prueba PPD tuvo un costo menor por individuo que la QFT-G (US$3,70 en comparación con US$18,60; P = 0.0008). CONCLUSIONES: La prueba QFT-G tuvo un costo más elevado, pero la tasa de detección fue más alta en la mayoría de los grupos destinatarios y el tiempo de respuesta fue más rápido en comparación con la PPD. Sin embargo, la sensibilidad de la prueba QFT-G fue inferior entre los individuos inmunodeficientes. Por consiguiente, se deben tomar las precauciones necesarias para utilizar la prueba QFT-G en el tamizaje y diagnóstico de ITBL en entornos de escasos recursos y alta prevalencia de VIH como Trinidad y Tabago.