The incidence of Orientia tsutsugamushi infection in rural South India.

Scrub typhus is a common bacterial infection in Asia caused by Orientia tsutsugamushi. This serological cohort study estimated the incidence of infection in a rural population in South India. Participants were enrolled through systematic sampling in 46 villages at baseline, and revisited the following year. Blood samples were tested for IgG antibodies using ELISA, followed by indirect immunofluorescence assays (IFA) in those positive for ELISA at both rounds. A case was defined as sero-conversion (ELISA), or at least a 4-fold titre increase (IFA), between the two time points. In addition to crude incidence rate estimates, we used piecewise linear rates across calendar months, with rates proportional to the monthly incidence of local hospital cases to address seasonality and unequal follow-up times. Of 402 participants, 61.7% were female. The mean age was 46.7 years, (range 13-88). 21 participants showed evidence for serological infection. The estimated incidence was 4.4 per 100 person-years (95% CI 2.8-6.7). The piecewise linear rates approach resulted in a similar estimate of 4.6 per 100 person years (95% CI 2.9-6.9). Considering previous estimates of symptomatic scrub typhus incidence in the same study population, only about 2-5% of infections may result in clinically relevant disease.

Clinical characteristics of and antibody response to spotted fever group rickettsial infections in South India: Case series and serological cohort study.

OBJECTIVE: The clinical and serological characteristics of spotted fever group rickettsial (SFGR) infections in South Asia are poorly understood. We studied the clinical presentation and the IgM/IgG response in cases enrolled at two health care centres in South India. METHOD: We enrolled 77 patients. Fifty-seven of these patients were recruited at a tertiary care centre, the remaining 20 at a community hospital (secondary care level). Diagnostic tests included IgM and IgG enzyme-linked immunosorbent assay and polymerase chain reaction. Over a period of 1 year, 41 cases were followed up for repeated sero-analysis. RESULTS: Median age was 9 years (range 1-79). A rash was present in 74% of cases (57/77). In cases aged <15 years, rash was present in 94% (44/47) vs. 43% (13/30) in cases aged ≥15 years. An eschar was found in two cases (3%). Severe infection or complications occurred in 10 cases (13%). These included central nervous system infection (6/77, 8%), kidney injury (3/77, 4%), shock (3/77, 4%), lung involvement (2/77, 3%) and peripheral gangrene (2/77, 3%). IgM antibody levels increased faster after fever onset than IgG antibodies, peaking at 50 and 60 days, respectively. After the peak, IgM and IgG levels showed a slow decline over one year with less than 50% of cases showing persistent IgG antibody levels. CONCLUSION: Spotted fever group rickettsial infections in South India may be under-diagnosed, as many cases may not develop a rash. The proportion of cases developing severe infection seems lower than for scrub typhus in this region. IgG seroprevalence may substantially underestimate the proportion in a population with past SFGR infection.

High initial IgG antibody levels against Orientia tsutsugamushi are associated with an increased risk of severe scrub typhus infection.

BACKGROUND: Scrub typhus is a dominant cause of febrile illness in many parts of Asia. Immunity is limited by the great strain diversity of Orientia tsutsugamushi. It is unclear whether previous infection protects from severe infection or enhances the risk. METHODS/PRINCIPAL FINDINGS: We studied IgG antibody levels against O. tsutsugamushi at presentation in 636 scrub typhus patients using enzyme-linked immunosorbent assays (ELISA). The association between ELISA optical density (OD) and risk of severe infection was modelled using Poisson regression. OD was categorised as low (<1.0), intermediate (1.0 to 2.9), and high (≥3.0). OD was also modelled as a continuous variable (cubic spline). Median age of cases was 41 years (range 0-85), with 37% having severe infection. Compared to the low category, the age-adjusted risk of severe infection was 1.5 times higher in the intermediate category (95%CI 1.2, 1.9), and 1.3 times higher in the high category (95%CI 1.0, 1.7). The effect was stronger in cases <40 years, doubling the risk in the intermediate and high categories compared to the low category. The effect was more pronounced in cases tested within 7 days of fever onset when IgG ODs are more likely to reflect pre-infection levels. CONCLUSIONS/SIGNIFICANCE: Intermediate and high IgG antibody levels at the time of diagnosis are associated with a higher risk of severe scrub typhus infection. The findings may be explained by severe infection eliciting an accelerated IgG response or by previous scrub typhus infection enhancing the severity of subsequent episodes.

Risk Factors for Scrub Typhus, Murine Typhus, and Spotted Fever Seropositivity in Urban Areas, Rural Plains, and Peri-Forest Hill Villages in South India: A Cross-Sectional Study.

Scrub typhus and spotted fever group rickettsioses are thought to be common causes of febrile illness in India, whereas they rarely test for murine typhus. This cross-sectional study explored the risk factors associated with scrub typhus, tick-borne spotted fever, and murine typhus seropositivity in three different geographical settings, urban, rural, and hill villages in Tamil Nadu, South India. We enrolled 1,353 participants living in 48 clusters. The study included a questionnaire survey and blood sampling. Blood was tested for Orientia tsutsugamushi (scrub typhus), Rickettsia typhi (murine typhus), and spotted fever group Rickettsia IgG using ELISA. The seroprevalence of scrub typhus, spotted fever, and murine typhus were 20.4%, 10.4%, and 5.4%, respectively. Scrub typhus had the highest prevalence in rural areas (28.1%), and spotted fever was most common in peri-forested areas (14.9%). Murine typhus was more common in rural (8.7%) than urban areas (5.4%) and absent in peri-forested hill areas. Agricultural workers had a higher relative risk for scrub typhus, especially in urban areas. For murine typhus, proximity to a waterbody and owning a dog were found to be major risk factors. The main risk factors for spotted fever were agricultural work and living in proximity to a forest. Urban, rural plains, and hill settings display distinct epidemiological pattern of Orientia and rickettsial infections. Although scrub typhus and spotted fever were associated with known risk factors in this study, the findings suggest a different ecology of murine typhus transmission compared with other studies conducted in Asia.

Antibody response following scrub typhus infection: clinical cohort study.

OBJECTIVE: Scrub typhus is a common cause of fever in Asia. The antibody response to infection and its effect on subsequent infection are unclear. We studied the IgM and IgG antibody response after infection, accounting for clinical severity. METHOD: We studied 197 scrub typhus patients for up to 2 years post-infection. Overall, 501 blood samples were analysed for scrub typhus antibodies using ELISA. IgM and IgG ELISA optical densities (OD) were analysed using quantile regression. OD values of 1.0 (IgM) and 1.5 (IgG) were used to define seropositivity. RESULTS: IgM OD values fell rapidly from an initial peak after infection. 50% of cases were IgM seronegative after 82 days. About 2 years after fever onset, 50% of cases had fitted IgG OD values of <1.5. Patients with high initial IgG OD values (≥2.5, used as a proxy for probable previous scrub typhus infection) had a more sustained IgG response than those with a low initial IgG OD, and more often presented with complications (18/36 = 50% vs. 28/91 = 30.8%, risk ratio = 1.63, 95% CI 1.04, 2.55, P = 0.035). This association was robust to adjusting for age (risk ratio 1.50, 95% CI 0.96, 2.33, P = 0.072). CONCLUSION: Cross-sectional IgG seroprevalence data substantially underestimate the proportion in a population ever infected with scrub typhus. A high initial IgG as a potential marker for previous scrub typhus infection may be associated with long-term IgG persistence and a higher risk of complicated scrub typhus.

OBJECTIF: Le typhus exfoliant est une cause fréquente de fièvre en Asie. La réponse anticorps à l'infection et son effet sur l'infection ultérieure ne sont pas clairs. Nous avons étudié la réponse des anticorps IgM et IgG après infection, en tenant compte de la sévérité clinique. MÉTHODE: Nous avons étudié 197 patients atteints de typhus exfoliant pendant une période allant jusqu'à deux ans après l'infection. Au total, 501 échantillons de sang ont été analysés pour déterminer la présence d'anticorps anti-typhus à l'aide d'ELISA. Les densités optiques (DO) des ELISA d'IgM et d'IgG ont été analysées par régression quantile. Les valeurs de DO de 1,0 (IgM) et 1,5 (IgG) ont été utilisées pour définir la séropositivité. RÉSULTATS: Les valeurs de DO d'IgM ont rapidement diminué par rapport au pic initial après infection. 50% des cas étaient séronégatifs pour les IgM après 82 jours. Environ 2 ans après le début de la fièvre, 50% des cas avaient des valeurs ajustées de OD d'IgG inférieures à 1,5. Les patients présentant des valeurs initiales de DO d'IgG élevées (≥2,5, utilisés comme proxy d'une infection antérieure probable par le typhus exfoliant) présentaient une réponse IgG plus maintenue que ceux présentant une DO initiale d'IgG faible et présentaient plus souvent des complications (18/36 = 50% vs. 28/91 = 30,8%, rapport de risque: 1,63; IC95%: 1,04-2,55, P = 0,035). Cette association était robuste pour l'ajustement en fonction de l'âge (rapport de risque: 1,50; IC95%: 0,96-2,33, P = 0,072). CONCLUSION: Les données transversales sur la séroprévalence des IgG sous-estiment considérablement la proportion dans une population déjà infectée par le typhus exfoliant. Des IgG initialement élevées en tant que marqueur potentiel d'infection antérieure par le typhus exfoliant peuvent être associées à une persistance à long terme des IgG et à un risque plus élevé de typhus exfoliant compliqué.

Hospitalisations and outpatient visits for undifferentiated fever attributable to scrub typhus in rural South India: Retrospective cohort and nested case-control study.

BACKGROUND: The burden of scrub typhus in endemic areas is poorly understood. This study aimed at estimating the proportion of hospitalisations and outpatient visits for undifferentiated fever in the community that may be attributable to scrub typhus. METHODOLOGY AND PRINCIPAL FINDINGS: The study was a retrospective cohort with a nested case-control study conducted in the South Indian state of Tamil Nadu. We conducted house-to-house screening in 48 villages (42965 people, 11964 households) to identify hospitalised or outpatient cases due to undifferentiated fever during the preceding scrub typhus season. We used scrub typhus IgG to determine past infection. We calculated adjusted odds ratios for the association between IgG positivity and case status. Odds ratios were used to estimate population attributable fractions (PAF) indicating the proportion of hospitalised and outpatient fever cases attributable to scrub typhus. We identified 58 cases of hospitalisation and 236 outpatient treatments. 562 people were enrolled as control group to estimate the background IgG sero-prevalence. IgG prevalence was 20.3% in controls, 26.3% in outpatient cases and 43.1% in hospitalised cases. The PAFs suggested that 29.5% of hospitalisations and 6.1% of outpatient cases may have been due to scrub typhus. In villages with a high IgG prevalence (defined as ≥15% among controls), the corresponding PAFs were 43.4% for hospitalisations and 5.6% for outpatients. The estimated annual incidence of scrub typhus was 0.8/1000 people (0.3/1000 in low, and 1.3/1000 in high prevalence villages). Evidence for recall error suggested that the true incidences may be about twice as high as these figures. CONCLUSIONS: The study suggests scrub typhus as an important cause for febrile hospitalisations in the community. The results confirm the adequacy of empirical treatment for scrub typhus in hospitalised cases with undifferentiated fever. Since scrub typhus may be rare among stable outpatients, the use of empirical treatment remains doubtful in these.