Evaluation of pooled ocular and vaginal swabs by the Cepheid GeneXpert CT/NG assay for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae compared to the GenProbe Aptima Combo 2 Assay.

Ocular swabs from Tanzania were tested for Chlamydia trachomatis (CT), and self-collected vaginal swabs collected through a home collection program, iwantthekit.org, were tested for Neisseria gonorrhoeae and CT to evaluate Cepheid GeneXpert for the use of pooling multiple specimens before testing. GeneXpert shows to be a promising test for pooling.

Trachoma and ocular Chlamydia trachomatis rates in children in trachoma-endemic communities enrolled for at least three years in the Tanzania National Trachoma Control Programme.

Trachoma, a blinding eye disease caused by repeated and prolonged infection with Chlamydia trachomatis, is a significant public health problem for sub-Saharan Africa. Tanzania has had a National Trachoma Task Force since 1999, working on trachoma control in endemic districts. The objective of this study was twofold: first, to determine the current status of infection and clinical trachoma in these districts in Tanzania, and second, to determine if a combination of clinical signs could be used as a surrogate for infection. We conducted a survey for trachoma and infection with C. trachomatis in 75 villages in eight districts of Kongwa, Kilosa, Mpwapwa, Bahi, Kondoa, Manyoni, Monduli and Iramba in Tanzania, which have previously been shown to be endemic. In each village, a random sample of households, and of children within households, was taken for examination. Trachoma was graded using the World Health Organization system, which we expanded, and a swab taken to determine presence of infection. The rates of trachoma ranged from 0% in Iramba District to 15.17% in Monduli District, with large variation in villages within districts. Infection rates were generally lower than trachoma rates, as expected, and most districts had villages with no infection. A combination of clinical signs of trachoma in children, when absent, showed very high specificity for identifying villages with no infection. We conclude that these signs might be useful for monitoring absence of infection in villages, and that districts with trachoma prevalence between 10% and 15% should have village level rapid surveys to avoid unnecessary mass treatment.

Gender equity and trichiasis surgery in the Vietnam and Tanzania national trachoma control programmes.

AIMS: To calculate the gender distribution of trichiasis cases in trachoma communities in Vietnam and Tanzania, and the gender distribution of surgical cases, to determine if women are using surgical services proportional to their needs. METHODS: Population based data from surveys done in Tanzania and Vietnam as part of the national trachoma control programmes were used to determine the rate of trichiasis by gender in the population. Surgical records provided data on the gender ratio of surgical cases. RESULTS: The rates of trichiasis in both countries are from 1.4-fold to sixfold higher in females compared to males. In both countries, the female to male rate of surgery was the same or even higher than the female to male rate of trichiasis in the population. CONCLUSIONS: These data provide assurance of gender equity in the provision and use of trichiasis surgery services in the national programmes of these two countries. Such simple analyses should be used by other programmes to assure gender equity in provision and use of trichiasis surgery services.

Prevalence of the different types of age-related cataract in an African population.

PURPOSE: To describe the prevalence of different types of cataract and their association with visual acuity in a Tanzanian population aged 40 years and older. METHODS: A prevalence survey for lens opacity, glaucoma, and visual impairment was carried out on all residents age 40 and older of six villages in Kongwa, Tanzania. One examiner graded the lens for presence of nuclear (NSC), posterior subcapsular (PSC), and cortical cataract (CC), using the new WHO Simplified Cataract Grading System. Visual acuity was measured in each eye, both presenting and best corrected, using an illiterate E chart. RESULTS: The proportion of eligible subjects participating was 90% (3268/3641). The prevalence of cataract was as follows: NSC, 15.6%; CC, 8.8%; and PSC, 1.9%. All types of cataract increased with age, from NSC, 1.7%; CC, 2.4%; and PSC, 0.4% for those aged 40 to 49 years to NSC, 59.2%; CC, 23.5%; and PSC, 5.9% for those aged 70 years and older (P < 0.0001 for all cataract types, chi(2) test for trend). Cataract prevalence was higher among women than men for NSC (P = 0.0001), but not for CC (P = 0.15) or PSC (P = 0.25), after adjusting for age. Prevalence rates of visual impairment (BCVA < 6/12), US blindness (< or = 6/60) and WHO blindness (< 6/120) for this population were 13.3%, 2.1%, and 1.3%, respectively. Older age and each of the major types of pure and mixed cataract were independently associated with worse vision in regression modeling. CONCLUSIONS: Unlike African-derived populations in Salisbury and Barbados, NSC rather than CC was most prevalent in this African population. The seeming lower prevalence of CC may to some extent be explained by different grading schemes, differential availability of cataract surgery, the younger mean age of the Tanzanian subjects, and a higher prevalence of NSC in this population.

OmpA genotypic evidence for persistent ocular Chlamydia trachomatis infection in Tanzanian village women.

BACKGROUND: Trachoma is still a significant problem in the developing world. Adult women are at higher risk of developing scarring and trichiasis, the potentially blinding sequelae, compared to men. In part, the higher risk may be due to more frequent infections in women because of their frequent contact with children, the main reservoir of C. trachomatis infection. However, other factors associated with infection, particularly constant infection, in adult women need to be identified. METHODS: A group of 118 women who were infected with C. trachomatis and 118 women who were not infected, but of similar age and trachoma status, were identified in 1996 from a population-based sample of women age 16 and older from eleven villages in Kongwa, Tanzania. This group of 236 was re-contacted three years later to ascertain trachoma status and determine infection status using polymerase chain reaction-enzyme immunoassay (PCR-EIA). Positive samples at both time points were examined for serovar and genotype shift, using ompA sequencing information. RESULTS: Of the original 236 women, 165 (70%) completed exams in 1999. Fifty-eight (35%) of the 165 women were excluded from this analysis because they received antibiotic treatment for trachoma in the six months prior to the second exam. Infection at baseline was the most important predictor of infection three years later (Age-adjusted odds ratio (95% confidence interval) 6.6 (1.8-24.4)). A total of 17 women (16%) were infected at the two examinations, and of the 15 for whom genotyping could be done, 11 (73%) were infected with the same ompA genotype at both time points. Chronically infected women were more likely to have trichiasis, scarring, and active trachoma at baseline than those never infected or infected only once. Only 41% of the chronically infected women were living in houses with infected pre-school children, but 24% were in houses with no children. Four of ten women with trichiasis developed incident corneal opacity over the three years. CONCLUSIONS: The data provide evidence for persistence of infection in a sub-group of women. The strongest predictor of infection at follow-up was baseline infection, and most were infected with the same genotype at both time points. For women with persistent infection, at least half were either not living with children or not living with infected children, suggesting that continual re-exposure from a close family member was less likely. Chronic infection is likely related to both exposure and immunological factors, and these need to be further identified. Inclusion of women in community-based treatment programs, regardless of whether a child is present in the house, is likely to be important in preventing the progression of inflammatory trachoma and scarring to trichiasis.

Progression of active trachoma to scarring in a cohort of Tanzanian children.

Risk factors for the incidence of scarring are needed to inform trachoma control programs in countries hyperendemic for this blinding disease. A cohort of pre-school children with constant, severe trachoma, and an age, sex, and neighborhood matched cohort of children without constant severe trachoma were followed for seven years to determine the incidence of scarring. The incidence of scarring in the children with constant severe trachoma was 29.2% versus 9.6% in the comparison group. In a model adjusting for multiple factors, significant predictors of scarring were increasing age, female, and constant severe trachoma (OR = 4.85, 95% CL = 2.05, 11.40). Infection with C. trachomatis at follow up was also associated with scarring in both groups of children. It is likely that these children have a different host response to infection, and represent a subgroup at high risk for the blinding complications of trachoma. Reducing exposure to infection in the community through antibiotics and changes in hygiene practices is still the most promising control strategy.

Azithromycin in control of trachoma.

BACKGROUND: Trachoma is the leading cause of preventable blindness. Programmes to prevent blindness due to trachoma are based on community-wide treatment with topical tetracycline. We assessed the potential of community-wide azithromycin treatment for trachoma control. METHODS: Pairs of villages in trachoma endemic areas of Egypt, The Gambia, and Tanzania were matched on trachoma rates in 1-10-year-old children. Villages were randomly assigned community-wide oral azithromycin treatment (three doses with intervals of 1 week) or treatment with 1% topical tetracycline (once daily for 6 weeks). Clinical examinations were done at baseline, 2-4.5 months, and 12-14 months after treatment. Chlamydia trachomatitis was identified by ligase chain reaction (LCR). Analyses were by intention to treat. Univariate comparisons and multivariate analyses were used to compare outcomes. FINDINGS: LCR positivity was correlated with clinical severity, but about 30% of Egyptian and Gambian villagers with no active disease were LCR positive. Village-wide LCR positivity ranged from 16.5% (Tanzania) to 43.6% (Egypt). Treatment compliance was over 90% except in the tetracycline treatment village in Egypt. Of the participants initially LCR positive, 866 (95%) of 924 who received at least one azithromycin dose and 482 (82%) of 587 who received 28 days or more topical tetracycline, were negative at follow-up. At 1 year, village-wide LCR positivity rates were substantially lower than at baseline with both treatments; the decreases were greater with azithromycin than with tetracycline (93% vs 77% in Egypt, 78 vs 66% in The Gambia, 64 vs 55% in Tanzania). Similarly, greater reduction in clinical activity occurred after azithromycin. In multivariate analyses, factors associated with being LCR positive at 1 year were: not receiving azithromycin; age under 10 years; and LCR positivity at baseline. INTERPRETATION: Community-wide treatment with oral azithromycin markedly reduces C. trachomatis infection and clinical trachoma in endemic areas and may be an important approach to control of trachoma.

Incidence of trichiasis in a cohort of women with and without scarring.

BACKGROUND: Blindness from trachoma is a significant problem for many underdeveloped countries. While active trachoma is common in children, trichiasis, the potentially blinding sequella, develops in adulthood and affects mainly women. Little is known about factors associated with the development of trichiasis. METHODS: The 7-year incidence of trichiasis and its association with ocular chlamydia infection was examined in a cohort of women from a hyperendemic area. A total of 4,932 women 18 years and older, living in 11 villages in Central Tanzania, were examined in 1989. A follow-up examination in 1996 was performed on all women with scars living in six of the 11 villages and on a random sample of women without scars from the same villages. Trachoma was graded clinically, chlamydia infection was ascertained at follow-up using polymerase chain reaction-enzyme immunoassay (PCR-EIA). RESULTS: A total 523 of the women with scars and 503 of the women without scars were re-examined. Forty-eight of the women with scars (incidence, 9.2%) and three of the women without scars (0.6%) developed trichiasis in the 7-year period. Prevalence of chlamydia infection was significantly higher in the group with scars (11.7% versus 7.1%). Trichiasis cases were more likely to be older, and to have chlamydia infection at follow-up odds ratio (95% confidence interval) 2.5 (1.1-5.7). CONCLUSION: The 7-year incidence rate in the population with scars was high, over 1% per year. Ocular chlamydia infection was more common in the group with scars at baseline and was also associated with being a trichiasis case, suggesting the importance of potentially long-term chlamydia infection in the progression to trichiasis. Antibiotic distribution programmes for trachoma control should include women with scars.

Evaluation of barriers to surgical compliance in the treatment of trichiasis.

PURPOSE: Eyelid repair surgery can prevent the effects of trichiasis leading to visual loss. Cost, transportation difficulties, and familial responsibilities have been identified as major barriers to surgical compliance. We evaluated whether offering trichiasis surgery in the village was effective in increasing the rate of surgical acceptance and in decreasing perceived barriers to surgery. METHODS: In 1989, 205 women with trichiasis were identified in Central Tanzania and were offered free surgery along with free transport. As of 1991, only 18% of these women had undergone the surgery. We followed-up these women 7 years later after village level surgery was introduced. RESULTS: Since 1991, an additional 12% of the women had undergone eyelid surgery. 44% were conducted in the village. Surgical cases since 1991 reported shorter travel times to the place of surgery, similar post-surgical problems, and fewer days in the hospital. While providing benefits to the patient, increased village eye services did not increase the rate of surgical acceptance. The women who declined surgery did not know surgery in the village was available and the perceived cost and transportation difficulties continued to be barriers. 50% of the non-acceptors stated that there was nothing that would enable them to accept surgical intervention despite the fact that 3/4 of them reported eye symptoms that interfered with their daily activities. CONCLUSIONS: The cost efficacy of village level eye services needs to be evaluated and the awareness of these services increased.

Evidence for a predominant proinflammatory conjunctival cytokine response in individuals with trachoma.

Immune responses to Chlamydia trachomatis infection in trachoma do not protect against reinfection or the development of scarring and blindness. In addition, the immunoregulatory contribution of cytokines to the development of conjunctival histopathology or protection is undefined. In this study, conjunctival cytokine mRNA transcripts were compared among subgroups of chlamydia infection status and ocular disease presentations of 50 individuals from an area where trachoma is endemic. There was a significant association of elevated interleukin (IL)-1beta, transforming growth factor beta1, and tumor necrosis factor alpha transcripts with infection, follicular inflammation, and scarring. Both gamma interferon (IFN-gamma) and IL-2 transcripts were significantly associated with infection; slightly elevated IL-2 levels were found in inflammatory disease. High IFN-gamma transcript levels were present with follicles and inflammatory disease and to a lesser extent with inflammatory scarring. The role of IFN-gamma in protection from infection or disease was not apparent from this study, since transcripts were frequently present in both chlamydial infection and disease. IL-12 (p40) transcripts were elevated in adults and children in association with follicular inflammation but not with scarring. IL-4, IL-5, and IL-10 transcripts were not detected in any samples. In conclusion, C. trachomatis infection stimulates local cytokines which favor a strong cell-mediated and proinflammatory response in both the early and later manifestations of trachoma. In addition, cytokine transcript levels that were associated with disease but no infection were characteristically lower overall than when chlamydia was present.

Nonocular Chlamydia infection and risk of ocular reinfection after mass treatment in a trachoma hyperendemic area.

PURPOSE: The presence of nasal discharge on a child's face increases the risk of active trachoma, suggesting that Chlamydia trachomatis in nasal secretions may be a possible source of ocular reinfection. The prevalence of chlamydia in nasal secretions and the risk of reinfection after mass treatment was investigated in a hyperendemic area of Tanzania. METHODS: In one village a total of 232 children aged 1 to 7 years were followed before and after mass treatment. Clinical trachoma, and microbiologic evidence of chlamydia, were assessed at baseline, 2 and 4 weeks into mass treatment, and 4 weeks after treatment stopped. The presence of chlamydia in ocular and nasal secretions was determined by polymerase chain reaction-enzyme immunoassay techniques. RESULTS: Of the 232 children, 59% had clinical trachoma and 27% had nasal specimens positive for chlamydia. Children with positive ocular chlamydia specimens and/or clinical trachoma were significantly more likely to have positive nasal specimens. At the end of mass treatment, only 4% of children had positive ocular specimens. However, 1 month after treatment stopped, the incidence of new infection was 21%. The rate of new ocular infections in those who had negative ocular specimens after treatment was similar between those who had positive and those who had negative nasal specimens at baseline. Positive ocular specimens at baseline was not a predictor of risk of new infection after treatment (odds ratio = 1.18, 95% confidence interval = 0.58, 2.40), suggesting these new infections were not the result of latent or persistent organism. CONCLUSIONS: These data do not support a role for nasal secretions in causing reinfection after treatment. One mass topical treatment alone is unlikely to be effective in trachoma hyperendemic areas as shown by the rapid re-emergence of infection.

The natural history of endemic trachoma: a longitudinal study.

A longitudinal study of trachoma was conducted among 100 members of nine families living in a hyperendemic area of Tanzania. Family members were examined for trachoma every three months for one year and conjunctival specimens were collected for antigen detection, which was performed either by direct fluorescent antibody cytologic analysis or enzyme immunoassay. The serovar specificity of tear antibodies was determined. Overall, young children tended to form a core of those with persistent, often severe, disease who consistently shed Chlamydia. These children are a potential source of infection in their family. Chlamydia could be identified early in the course of presumed recently acquired infections, but not later in resolving infections. This temporal change may account for the discrepancies between demonstrable organisms and clinical disease seen in cross-sectional studies. Several children were identified who did not develop trachoma despite having Chlamydia identified in conjunctival scrapings. Their ability to resist infection may offer clues for vaccine development. The study of serovar specificity is consistent with the intrafamily transmission of trachoma, but was confounded by the large family size and the potential for separate transmission units to occur within large extended families. These observations give further understanding of the natural history and kinetics of the transmission of trachoma that should be of use in developing and evaluating intervention studies.

Diagnosis of Chlamydia trachomatis eye infection in Tanzania by polymerase chain reaction/enzyme immunoassay.

Detection of Chlamydia trachomatis eye infection is largely unsatisfactory by standard laboratory methods. A polymerase chain reaction/enzyme immunoassay (PCR-EIA) that had previously been successful for diagnosis of genital C trachomatis infection was compared with direct antibody immunofluorescence (DFA) for detection of the organism in conjunctival scrapes. 234 Tanzanian children aged 1-7 years living in a village that had had no previous trachoma control programme were classified clinically as having no sign of trachoma (0) n = 97, follicular trachoma (TF) n = 100, or intense inflammatory trachoma with or without TF (TI +/- TF) n = 37. PCR-EIA detected C trachomatis in 24%, 54%, and 95% of subjects, respectively, compared with elementary body (EB) detection by DFA of 1%, 28%, and 60%, respectively. Overall prevalence of chlamydial eye infection was 22% by DFA compared with 48% by PCR-EIA. Of subjects with chlamydial DNA at pretreatment, 103 (92%) had no detectable chlamydial DNA at the end of 4 weeks of ocular tetracycline. The findings show that PCR-EIA is likely to affect trachoma diagnosis and epidemiology because of the increased sensitivity for detection of C trachomatis in all clinical groups; the less stringent requirements for specimen collection and transport make this method suitable for field use. Moreover, the semi-quantitative aspect of PCR-EIA may be useful for monitoring a decrease in chlamydial DNA after treatment.

Longitudinal study of the microbiology of endemic trachoma.

A longitudinal study of trachoma in 100 members of nine Tanzanian families was conducted to assess the sources of variation in the laboratory identification of trachoma and the changes that might occur over time. Multiple conjunctival swabs were collected every 3 months for 1 year and examined by direct fluorescent-antibody cytology (DFA), enzyme immunoassay, or microimmunofluorescence serology for tear antichlamydial antibodies. DFA specimens collected 5 min apart had a discordance rate of 10% and this is attributable to sampling variation. DFA specimens collected 2 days or more apart show a 25% discordance rate. This suggests a biologic variation in shedding in addition to sampling variation. Good correlation existed between the DFA and the enzyme immunoassay. Tear serology was quite specific in predicting the presence of clinical disease and correlated with the other two antigen detection tests, although it does not seem to offer any practical advantages. These studies indicate that there is considerable variation in the shedding of chlamydia by people living in trachoma-endemic areas.