TST indicates tuberculin skin test; IFN-γ, interferon γ. TST
cutpoint is at least 10 mm; IFN-γ assay cutpoint is at least 0.35 IU/mL.
TST indicates tuberculin skin test; IFN-γ, interferon γ. The
overlaid curve is a plot of the probability density function, a smoothed version
of the histogram.
TST indicates tuberculin skin test; IFN-γ, interferon γ. The
boxes, which indicate the distribution of IFN-γ values for each category
of TST induration, are bordered at the 25th and 75th percentiles of the IFN-γ
values, with a median line at the 50th percentile. Outliers are not displayed.
IFN-γ values of more than 10 IU/mL are displayed as 10 IU/mL. The horizontal
dotted line indicates the IFN-γ assay cutpoint of 0.35 IU/mL.
TST indicates tuberculin skin test; IFN-γ, interferon γ. The
overlaid curves are plots of the probability density functions, smoothed versions
of the histograms.
Pai M, Gokhale K, Joshi R, Dogra S, Kalantri S, Mendiratta DK, Narang P, Daley CL, Granich RM, Mazurek GH, Reingold AL, Riley LW, Colford JM. Mycobacterium tuberculosis Infection in Health Care Workers in Rural IndiaComparison of a Whole-Blood Interferon γ Assay With Tuberculin Skin Testing. JAMA. 2005;293(22):2746-2755. doi:10.1001/jama.293.22.2746
Author Affiliations: Departments of Medicine
and Microbiology, Mahatma Gandhi Institute of Medical Sciences, Sevagram,
India (Drs Pai, Gokhale, Joshi, Dogra, Kalantri, Mendiratta, and Narang);
Divisions of Epidemiology and Infectious Diseases, School of Public Health,
University of California, Berkeley (Drs Pai, Reingold, Riley, and Colford);
Department of Medicine, University of California, San Francisco (Dr Daley);
Division of Tuberculosis Elimination, Centers for Disease Control and Prevention,
Atlanta, Ga (Drs Granich and Mazurek); and Office of World Health Organization
Representative to India, New Delhi (Dr Granich). Dr Daley is now with the
National Jewish Medical and Research Center, Denver, Colo. Dr Granich is now
with the Office of the United States Global AIDS Coordinator, Washington,
Context Mycobacterium tuberculosis infection in health
care workers has not been adequately studied in developing countries using
newer diagnostic tests.
Objectives To estimate latent tuberculosis infection prevalence in health care
workers using the tuberculin skin test (TST) and a whole-blood interferon γ
(IFN-γ) assay; to determine agreement between the tests; and to compare
their correlation with risk factors.
Design, Setting, and Participants A cross-sectional comparison study of 726 health care workers aged 18
to 61 years (median age, 22 years) with no history of active tuberculosis
conducted from January to May 2004, at a rural medical school in India. A
total of 493 (68%) of the health care workers had direct contact with patients
with tuberculosis and 514 (71%) had BCG vaccine scars.
Interventions Tuberculin skin testing was performed using 1-TU dose of purified protein
derivative RT23, and the IFN-γ assay was performed by measuring IFN-γ
response to early secreted antigenic target 6, culture filtrate protein 10,
and a portion of tuberculosis antigen TB7.7.
Main Outcome Measures Agreement between TST and the IFN-γ assay, and comparison of the
tests with respect to their association with risk factors.
Results A large proportion of the health care workers were latently infected;
360 (50%) were positive by either TST or IFN-γ assay, and 226 (31%) were
positive by both tests. The prevalence estimates of TST and IFN-γ assay
positivity were comparable (41%; 95% confidence interval [CI], 38%-45% and
40%; 95% CI, 37%-43%, respectively). Agreement between the tests was high
(81.4%; κ = 0.61; 95% CI, 0.56-0.67). Increasing age and years
in the health profession were significant risk factors for both IFN-γ
assay and TST positivity. BCG vaccination had little impact on TST and IFN-γ
Conclusions Our study showed high latent tuberculosis infection prevalence in Indian
health care workers, high agreement between TST and IFN-γ assay, and similar
association between positive test results and risk factors. Although TST and
IFN-γ assay appear comparable in this population, they have different
performance and operational characteristics; therefore, the decision to select
one test over the other will depend on the population, purpose of testing,
and resource availability.
An estimated one third of the world’s population is infected with Mycobacterium tuberculosis,1 presenting
a major impediment to tuberculosis control. Despite the importance of latent
tuberculosis infection (LTBI), the tuberculin skin test (TST) was, until recently,
the only test available for its diagnosis.2- 5 The
TST measures hypersensitivity response to purified protein derivative (PPD),
a crude mixture of antigens, many of which are shared among M tuberculosis, Mycobacterium bovis bacille
Calmette-Guérin (BCG), and several nontuberculous mycobacteria (NTM).
The TST has limitations with respect to accuracy and reliability.2- 6
Advances in genomics and immunology have led to a promising alternative,
the in vitro interferon γ (IFN-γ) assay,6- 8 based
on the concept that T cells of infected individuals release IFN-γ. Although
early IFN-γ assays used PPD, newer assays use antigens, such as the
early secreted antigenic target 6 (ESAT-6) and culture filtrate protein 10
(CFP-10). These proteins, encoded within the region of difference 1 (RD1)
of the M tuberculosis genome, are significantly more
specific to M tuberculosis than PPD, as they are
not shared with BCG substrains or several NTM species that might cause nonspecific
sensitization.6 The QuantiFERON-TB assay (Cellestis
Limited, Carnegie, Australia) was the first commercially available IFN-γ
assay. The PPD-based QuantiFERON-TB test was approved by the US Food and Drug
Administration in 2001.9 The QuantiFERON-TB
Gold, an enhanced assay based on RD1 antigens, was approved by the Food and
Drug Administration in December 2004. A simplified variant of the QuantiFERON-TB
Gold assay, the QuantiFERON-TB Gold In-Tube test (hereafter referred to as
IFN-γ assay) has been recently developed. This assay uses overlapping
peptides representing ESAT-6, CFP-10, and a portion of tuberculosis antigen
TB7.7 (Rv2654), and stimulation by this antigenic mixture occurs within the
tube used to collect blood. Other tests, including those using enzyme-linked
immunospot, have also been evaluated.6- 8
Several studies have evaluated IFN-γ assays (in a review article8); however, very few have been conducted in tuberculosis-endemic
countries. In such countries, high BCG vaccine coverage, widespread NTM exposure,
and high tuberculosis incidence pose challenges for the evaluation of any
LTBI test. Because of the paucity of data from endemic countries and the need
for improved tuberculosis diagnostic tests, we evaluated an RD1-based IFN-γ
assay in a high-risk population in India, the country that accounts for one
third of the global burden of tuberculosis.10,11 An
estimated 40% of the Indian population is infected and the annual risk of
infection is 1.5%.10- 12
Health care workers are at high risk for tuberculosis.13 Despite
India’s tuberculosis burden, little is known about tuberculosis in health
care workers.14,15 The Revised
National Tuberculosis Control Programme of India is focused on expanding Directly
Observed Treatment, Short-Course services. There are currently no national
guidelines specific for reducing nosocomial tuberculosis risk. Because health
care workers treat large numbers of patients16 and
isolation facilities are scarce and expensive, Indian hospitals do not routinely
isolate patients with tuberculosis.14,15 Health
care workers often work in crowded hospitals with minimal infection control
measures.14 The goals of our study were to
estimate the burden of LTBI in Indian health care workers using the TST and
an enhanced IFN-γ assay, determine the agreement between the tests, and
compare their correlation with LTBI risk factors.
We conducted a cross-sectional comparison study from January to May
2004, at the Mahatma Gandhi Institute of Medical Sciences, a rural medical
school in Sevagram, India. At this hospital, about 300 patients with tuberculosis
are treated each year and the estimated community prevalence of smear-positive
tuberculosis is 200 per 100 000,17 with
the annual risk of infection at 1.2% to 1.6%.18 At
the Mahatma Gandhi Institute of Medical Sciences hospital, there are limited
tuberculosis infection control measures in place and health care workers are
constantly exposed to patients with tuberculosis. Although no data were available
on the incidence of tuberculosis among health care workers at the time of
our study, 5 (33%) of 15 internal medicine residents had developed active
tuberculosis during their training.
Our study was designed to recruit health care workers, including trainees,
nurses, laboratory workers, orderlies, and attending physicians. Health care
workers who met any of the following criteria were excluded: age younger than
18 years, pregnancy, allergy to tuberculin, and active tuberculosis in the
past. Because our goal was to recruit all health care workers at our institution,
no formal sample size determination was made. All participants gave written
informed consent, and the research was approved by the ethics committees at
the Mahatma Gandhi Institute of Medical Sciences hospital and the University
of California, Berkeley.
Each participant provided data on age, sex, education, employment, BCG
vaccination, prior TST, and symptoms. We also assessed surrogate markers of M tuberculosis exposure, including year of training, years
in the health care profession, job category, direct contact with sputum smear-positive
tuberculosis (defined as contact between 2 people that is of sufficient distance
to allow conversation between them5), training
in internal medicine, and household tuberculosis contact. Education and job
category were used as socioeconomic status indicators. After the interview,
BCG vaccine scar was ascertained by inspection and blood was drawn for the
IFN-γ assay, followed by TST administration. The tests were interpreted
independently of each other and of exposure information. Human immunodeficiency
virus testing was not performed because of the low prevalence of positive
human immunodeficiency virus expected in this young, educated population.
Participants who were symptomatic or positive by either test were investigated
(using chest radiographs and sputum microscopy) to rule out active disease.
Although it is not standard practice to treat LTBI in India, health care workers
diagnosed to have LTBI (n = 360) in our study were offered the option
of counseling on risks and benefits of LTBI treatment. Of these, 61 (17%)
health care workers (mostly student and staff nurses) elected to undergo standard
isoniazid treatment for 6 months.19 As of January
2005, 37 (61%) of 61 health care workers had completed therapy with no major
As recommended by India’s Revised National Tuberculosis Control
Programme20 and based on previous studies,3,21 we performed the TST using 1-TU dose
of PPD RT23 (Statens Serum Institut, Copenhagen, Denmark). Tuberculin skin
test was administered by a certified technician using the Mantoux method and
read after 48 to 72 hours using a blinded caliper. Interreader variability
was assessed using a second independent reader for a subset of the participants.
The second trained reader used the same palpation technique as the first.
As recommended by the Revised National Tuberculosis Control Programme20 and consistent with the standard risk-stratified
interpretation,19 an induration of at least
10 mm was considered positive. For comparison, 5 mm and 15 mm cutpoints were
An enhanced “in-tube” version of the IFN-γ assay was
used for this study. The assay involves 2 stages: incubation of whole blood
with antigens, and measurement of IFN-γ production in harvested plasma
by enzyme-linked immunosorbent assay. Venous blood was directly collected
into three 1-mL heparin-containing tubes. One tube contained only heparin
as negative control, another also contained the T-cell mitogen phytohemagglutinin
as positive control, and the third tube had overlapping peptides representing
the entire sequences of ESAT-6 and CFP-10, and another peptide representing
a portion of TB7.7. Within 2 to 6 hours of blood draw, the tubes were incubated
at 37°C. After 24 hours of incubation, the tubes were centrifuged and
plasma was harvested and frozen at –70°C until the enzyme-linked
immunosorbent assay was performed. The amount of IFN-γ was quantified
using the enzyme-linked immunosorbent assay. The IFN-γ values (IU/mL)
for tuberculosis-specific antigens and mitogen were corrected for background
by subtracting the value obtained for the respective negative control. As
recommended by the manufacturer and based on previous studies,22,23 the
cutoff value for a positive test was IFN-γ of at least 0.35 IU/mL. The
enzyme-linked immunosorbent assay cannot accurately determine the absolute
IFN-γ values when they exceed 10 IU/mL. Therefore, IFN-γ values
of more than 10 IU/mL are shown as 10 IU/mL in our graphs. Testing was performed
at the study site, and all assays met quality control standards.
Statistical analyses were performed using Stata version 8 (Stata Corp,
College Station, Tex). The outcomes included agreement between the tests,
factors associated with discordance, and comparison of the tests with respect
to their association with risk factors. Agreement between the tests was quantified
using the κ statistic. Correlation was also assessed between continuous
IFN-γ values (IU/mL) and TST induration (mm). Factors associated with
discordance were explored using a logistic regression model with any discordance
(either positive TST/negative IFN-γ assay or negative TST/positive IFN-γ
assay) as the outcome, and factors such as age, sex, exposure to tuberculosis,
and BCG vaccine as covariates.
Risk factors for test positivity were evaluated using odds ratios (ORs).
To adjust for multiple covariates, we used a logistic regression model with
TST or IFN-γ assay results as the outcome and covariates such as markers
of tuberculosis exposure. Logistic regression was performed using methods
described elsewhere24: covariates that were
significant in bivariate analyses were included in the preliminary model.
In addition, other covariates (such as age) that were considered biologically
important were forced into the model irrespective of statistical significance.
Interaction between covariates was evaluated by adding interaction terms to
the model and evaluating its statistical significance using the Wald P value. From the initial model, covariates that did not
contribute significantly were dropped 1 at a time until the best fitting,
parsimonious model was identified. Model fit was evaluated using the Hosmer-Lemeshow
test.24 The results of the final model are
presented as adjusted ORs with 95% confidence intervals (CIs). Because young
trainees in the hospital were expected to have shorter and uncomplicated exposure
histories, and recall of exposure might be better in students, we planned
a prespecified subgroup analysis to evaluate risk factors among 353 medical
and nursing students.
Of 1081 eligible health care workers, 726 (67%) participated in the
study (Figure 1). Participation rates
were high among students (91%) but low among attending physicians (10%). As
shown in Table 1, medical and nursing
students accounted for 48% of the cohort. The median age was 22 years (range,
18-61 years). Only 36 of the health care workers (5%) had ever been tuberculin
tested and 514 (71%) of 726 had BCG vaccine scars. A total of 493 (68%) of
the participants reported direct contact with patients with sputum smear-positive
tuberculosis. Exposure was particularly high among interns, residents, physicians,
and nurses. On average, these health care workers reported at least 3 contacts
with patients with tuberculosis in the week before their interviews.
Valid TST results were available in 720 (99%) of 726 individuals. Figure 2 shows a bimodal TST distribution with
the first peak centered on zero and a second peak centered on 15. With a cutpoint
of at least 10 mm, 298 (41%) of 720 participants were positive (95% CI, 38%-45%).
With a cutpoint of at least 15 mm, 167 (23%) of 720 participants were positive
(95% CI, 20%-27%). A second TST reader independently read 254 (35%) of 720
participants and the interrater agreement was excellent (κ = 0.91). Table 2 shows the risk factors for TST positivity.
On multivariate analysis, age (21-30 vs 18-20 years) and number of years in
the health care profession (>5 vs ≤1 year) were statistically significant.
Previous BCG vaccination was not associated with TST positivity in bivariate
and multivariate analyses.
Valid IFN-γ assay results were available for 725 (99%) of 726 participants
(Figure 2). The median IFN-γ assay
level was 0.11 IU/mL. With a cutpoint of at least 0.35 IU/mL, 291 (40%) of
725 participants were positive (95% CI, 37%-43%). Table 2 shows the risk factors for IFN-γ assay positivity. On
multivariate analysis, age (≥41 vs 18-20 years), number of years in the
health profession (>5 vs ≤1 year), and job category (orderlies vs medical
students) were significant. Previous BCG vaccination was not associated with
IFN-γ assay positivity in bivariate and multivariate analyses.
Among students, we evaluated covariates, including age, sex, student
category, socioeconomic status, year of training, direct contact with active
tuberculosis, household contact, training in internal medicine, and BCG vaccine
scar. With TST as the outcome, the final model had only 2 covariates: age
and sex. Students aged 21 to 23 years and older than 23 years had increased
ORs compared with students aged 18 to 20 years (1.45; 95% CI, 0.8-2.6; and
4.2; 95% CI, 1.9-9.3; respectively). Although not statistically significant,
female students had higher odds of TST positivity (OR, 1.7; 95% CI, 0.9-2.9).
With IFN-γ assay as the outcome, only age and sex were retained in the
final model. The ORs for those participants aged 21 to 23 years and older
than 23 years were 1.36 (95% CI, 0.8-2.4) and 4.01 (95% CI, 1.8-8.8), respectively.
Female students had higher odds of IFN-γ assay positivity (OR, 1.6; 95%
CI, 0.9-2.8) but was not statistically significant.
Data on agreement between the TST and IFN-γ assay results were available
for 719 participants (Table 3). Of these,
360 (50%) were positive by either test and 226 (31%) were positive by both
tests. With a 10-mm cutpoint, the agreement was 81.4% (κ = 0.61;
95% CI, 0.56-0.67). Agreement was lower when 5-mm and 15-mm cutpoints were
used. Figure 3 shows the distribution
of IFN-γ assay values by TST category. A sharp increase in the IFN-γ
assay values was observed when the induration exceeded 10 mm. Figure 4 shows the TST distribution by IFN-γ assay status. The
TST distributions are well-separated, with a median of 3 mm and 15 mm in the
negative IFN-γ assay and positive IFN-γ assay groups, respectively
(P<.001 for difference between medians, using
the Wilcoxon rank sum test).
Table 4 shows the agreement data
stratified by covariates. Increasing age, years in health profession, lower
educational level, and specific job categories (laboratory staff, orderlies,
and attending physicians/faculty) were associated with greater discordance
(either positive TST/negative IFN-γ assay or negative TST/postive IFN-γ
assay) on bivariate analyses. However, on multivariate analysis, only 2 covariates
were important but not statistically significant: job category (attending
physicians/faculty vs medical students: OR, 3.9; 95% CI, 0.9-15.6) and increasing
years in health care (OR, 1.3; 95% CI, 0.6-2.8; 2.01; 95% CI, 0.8-5.4; and
2.1; 95% CI, 0.6-7.5; respectively) for those participants with 2 to 5 years,
6 to 10 years, and more than 10 years in the health profession compared with
those with 1 year or less. A positive TST/negative IFN-γ assay pattern
of discordance was more frequently observed with increasing age, increasing
years in the health care profession, and in specific subgroups (nurses, laboratory
staff, and attending physicians/faculty). A negative TST/positive IFN-γ
assay pattern of discordance was more frequently observed in those participants
with household tuberculosis contact. However, none of these differences were
statistically significant on multivariate analyses (data not shown).
Our study demonstrated the expected high prevalence of LTBI in Indian
health care workers. Although TST and IFN-γ assay use different antigen
combinations, it was surprising that these tests had comparable prevalence
estimates (41% and 40%, respectively) and a high level of agreement (81.4%, κ = 0.61).
The marginal bimodal TST distribution appears to be composed of 2 fairly well-separated
component distributions of negative and positive IFN-γ assay individuals.
Greater discordance between the PPD-based TST and the tuberculosis-specific
antigen–based IFN-γ assay was expected. The use of 1-TU dosage of
PPD in our study may have improved TST specificity, as many countries use
a 2-TU dosage.21 Increasing age and years in
the health profession were risk factors for both IFN-γ assay and TST positivity,
and the risk factor associations were fairly similar for both tests. To our
knowledge, our study is the largest evaluation of an IFN-γ assay among
health care workers in a developing country and the first report of the In-Tube
version of the QuantiFERON-TB Gold assay. This version of the assay is easier
to perform and facilitates testing of people in remote locations.
The high prevalence of LTBI in our study is consistent with estimates
from other developing countries.13,25- 27 However,
our prevalence of approximately 40% might be an underestimate because we recruited
few senior physicians who were older and, presumably, more frequently exposed
to M tuberculosis. Also, our study was not designed
to determine if tuberculosis transmission occurred in the nosocomial setting.
Although studies from India have suggested a high rate of tuberculosis among
health care workers,14,15 these
studies did not test for LTBI.
The risk factors we identified have been shown to be important in several
studies.13,25- 29 Age
and years in health care reflect cumulative exposure to M tuberculosis. Variability of risk across job categories may reflect
variations in exposure frequency and intensity.28,29 Although
direct contact with patients with tuberculosis has been shown to be a risk
factor in previous studies,13,25- 27 we
did not find a strong effect. However, this is not surprising as nearly 70%
of our health care workers reported direct contact, reducing the predictive
ability of this exposure factor. Overall, our findings highlight the need
to study tuberculosis among Indian health care workers. India’s Revised
National Tuberculosis Control Programme is the largest Directly Observed Treatment,
Short-Course program in the world,16 and the
safety and well-being of health care workers is important for the continued
expansion of this successful program. Further research is needed to document
the incidence of tuberculosis among Indian health care workers and determine
the rate of nosocomial transmission. If such studies demonstrate a problem,
control strategies will be needed to address the issue.
Several studies have compared RD1-based IFN-γ assays with TST,
mostly in low-endemic populations.8 The agreement
in our high-endemic population (81.4%), interestingly, is consistent with
those data reported by a majority of the studies.22,30- 33 A
recent study on an RD1-based IFN-γ assay that reported agreement data
showed an agreement of 94%, but this analysis included participants who had
not received BCG vaccine.22 Only 1 published
study has estimated the prevalence of LTBI in India using an IFN-γ assay.
Lalvani et al34 showed that 80% of healthy
adults (affluent corporate executives) in Bombay (Mumbai) tested enzyme-linked
immunospot−positive to either ESAT-6 or CFP-10. In contrast, only 40%
of our high-risk cohort was positive by IFN-γ assay, in line with the
epidemiological estimate for India of 40%10 and
the 41% TST positivity in the present study. Differences in IFN-γ assay
methodology might explain this difference.
Although previous studies have shown BCG vaccine to cause a positive
TST/negative IFN-γ assay discordance,31,35 previous
BCG vaccination had little impact on TST in our study. Our study largely comprised
adults who received BCG vaccine (Danish 1331 strain) at birth. In individuals
vaccinated in infancy, TST response due to BCG vaccine wanes rapidly.3,5,36,37 Another
explanation could be misclassification of BCG vaccine status owing to the
use of scar as a proxy.3,36 Recent
tuberculin surveys from India, involving more than 100 000 children,
have also shown the BCG vaccination does not influence the estimation of annual
risk of infection.36,38,39 However,
BCG vaccine might have an effect on TST in other populations, depending on
vaccine strain, timing, frequency, and time since vaccination.4,5,37 Infection
with NTM, which tends to cause nonspecific tuberculin sensitivity, is highly
prevalent in India.3,40 Therefore,
NTM infection might have caused false-positive TST results. Because the IFN-γ
assay uses RD1 antigens, the effect of NTM on the test should be limited.
However, ESAT-6 and CFP-10 have the potential to cross-react with certain
NTM species,6,30 and it has been
suggested that cross-reactivity may exist with Mycobacterium
leprae.41 Because NTM and M leprae are endemic in India, research is needed to study the effect
of NTM on IFN-γ assays.
Our cross-sectional data suggest that both tests are comparable in the
population we studied. However, this does not necessarily indicate that these
tests are completely equivalent because they do not measure exactly the same
immunological phenomenon.35 To put our findings
in context, it is important to review the evidence on the performance characteristics
of these tests.
The TST has moderate to high sensitivity and specificity, depending
on the population screened, with specificity being more unpredictable.2- 5 Several
studies have shown a positive association between TST response and subsequent
risk of active tuberculosis,2,4,42,43 and
randomized trials have shown that treatment of LTBI, diagnosed using TST,
reduces the risk of active tuberculosis by 60% to 90%.19 Furthermore,
the TST is a simple test with low material costs that does not require a laboratory,
but does require skilled testers.
The RD1-based IFN-γ assays have higher specificity than TST, better
correlation with exposure to M tuberculosis than
TST, no cross-reactivity due to BCG vaccination, and limited cross-reactivity
due to NTM infection.6- 9,22,23,31,35 Other
advantages include rapidity, the need for only a single visit, avoidance of
subjective measurements, and boosting.6,7,35 There
is limited evidence of an association between IFN-γ assay response to
ESAT-6 and subsequent progression to active tuberculosis among contacts of
patients with tuberculosis.44 To date, no trial
has demonstrated the efficacy of treatment based on IFN-γ assay results.
A limitation of IFN-γ assays is its higher material costs and the need
for laboratory infrastructure.8 Also, IFN-γ
assays have been inadequately evaluated in children, individuals with immunosuppressive
conditions, and in high-burden countries.8
Our study has several limitations. Unlike the previous IFN-γ assay
studies in outbreak and contact investigations,22,31- 33 we
were unable to quantify duration and proximity of contact with patients with
tuberculosis. Such exposure measures might have enabled us to evaluate whether
the IFN-γ assay was better associated with increasing exposure than TST,
although in a high prevalence health care worker population this may be difficult.
Because of the cross-sectional design and the lack of follow-up data, we were
unable to adequately resolve the discordance between the 2 tests.
We were also unable to measure important covariates, such as NTM and
human immunodeficiency virus infection. Information on human immunodeficiency
virus status would have helped to better evaluate the performance of the IFN-γ
assay. Although nonresponse among senior physicians might have led to an underestimation
of LTBI prevalence, it is unlikely to affect the comparisons between TST and
the IFN-γ assay. Unfortunately, due to lack of data from nonparticipants,
we could not compare their characteristics with those who did participate.
Although consistent with the Indian policy, the use of 1-TU dose of
PPD RT23 limits our ability to compare our data with studies that have used
2-TU dosages. Our study consisted of health care workers in a high prevalence
country. Although this may limit our ability to generalize the results to
settings with different baseline prevalences, we believe our data will be
helpful in understanding the performance of IFN-γ assays in high-burden
settings for which data are scarce.8
In conclusion, our study showed a high prevalence of LTBI in Indian
health care workers and high agreement between TST and an RD1-based IFN-γ
assay. Our results and the available evidence suggest that both tests have
advantages and limitations and, as of this time, both may have a useful role,
depending on factors unique to each setting. The emergence of IFN-γ
assays is a welcome development that has expanded the armamentarium of LTBI
diagnostics. The decision to select one or the other test will depend on the
population, the goal of testing, and the resources available. For example,
for serial testing of health care workers, the IFN-γ assay will be appropriate.
It will eliminate the need for repeat visits, avoid boosting, and minimize
interpretational difficulties. Because of its high specificity, RD1-based
assays will be helpful in populations where cross-reactivity due to BCG vaccine
and NTM pose problems in TST interpretation and/or where true tuberculosis
infection rate is relatively low. Furthermore, the IFN-γ assay may be
helpful in screening populations in which low return rates for TST reading
are a concern.
On the other hand, in high-burden, resource-limited settings, the TST
might serve a useful purpose. In India, the TST is widely used in diagnosing
childhood tuberculosis,3,20 and
in epidemiological studies on annual risk of infection in which blood testing
will be difficult.3,10,12,18,21,36 In
India, a 15-year follow-up of 280 000 individuals showed that TST response
is significantly associated with development of active tuberculosis.42 Our findings and evidence from Indian studies3,10,18,21,36 suggest
that the TST remains a useful test, particularly because of the low cost and
since BCG vaccine has a limited effect on TST.36,38,39 The
relationship between BCG vaccine and subsequent TST response may vary in other
populations. To fully evaluate the use of IFN-γ assays, long-term cohort
studies to determine the association between positive IFN-γ assay results
and the subsequent risk of active tuberculosis are required in diverse settings.8 If such studies demonstrate a strong consistent association,
IFN-γ assays might have the potential to replace TST.
Corresponding Author: Madhukar Pai, MD,
PhD, Division of Epidemiology, University of California, Berkeley, 140 Warren
Hall, Berkeley, CA 94720 (firstname.lastname@example.org).
Author Contributions: Dr Pai had full access
to all of the data in the study and takes responsibility for the integrity
of the data and the accuracy of the data analysis.
Study concept and design: Pai, Kalantri, Narang,
Daley, Granich, Mazurek, Riley, Colford.
Acquisition of data: Pai, Gokhale, Joshi, Dogra,
Kalantri, Mendiratta, Granich.
Analysis and interpretation of data: Pai, Joshi,
Kalantri, Narang, Granich, Mazurek, Reingold, Riley, Colford.
Drafting of the manuscript: Pai, Kalantri,
Critical revision of the manuscript for important
intellectual content: Pai, Gokhale, Joshi, Dogra, Kalantri, Mendiratta,
Narang, Daley, Granich, Mazurek, Reingold, Riley, Colford.
Statistical analysis: Pai, Colford.
Obtained funding: Pai, Reingold.
Administrative, technical, or material support:
Gokhale, Joshi, Dogra, Mendiratta, Narang, Granich, Mazurek, Reingold, Riley,
Study supervision: Pai, Joshi, Kalantri, Mendiratta,
Daley, Granich, Reingold, Riley, Colford.
Financial Disclosures: None of the investigators
have any financial interest in or a financial conflict with the subject matter
or materials discussed in this manuscript. Patent protection for incubation
of whole blood and antigens within the collection tube that contains a gel
plug for subsequent plasma separation and storage may be requested by the
US Centers for Disease Control and Prevention, listing Gerald Mazurek, MD,
as one of the inventors.
Funding/Support: Dr Pai was supported by a
doctoral training fellowship from the Fogarty AIDS International Training
and Research Program (grant 1-D43-TW00003-16). QuantiFERON-TB Gold diagnostic
kits were provided by Cellestis Limited, Carnegie, Australia.
Role of the Sponsor: The Fogarty AIDS International
Training and Research Program had no role in the study design, conduct, collection,
management, analysis, or interpretation of the data, or preparation, review,
or approval of the manuscript. The Cellestis Limited Chief Scientific Officer
provided input in study design and laboratory aspects of the QuantiFERON-TB
Gold assay. This company did not provide funding for this study and had no
role in the conduct, collection, management, analysis, or interpretation of
the data, or preparation, review, or approval of the manuscript.
Previous Presentation: Presented in part at
the Annual International Conference of the American Thoracic Society, San
Diego, Calif, May 22, 2005.
Acknowledgment: We thank all the health care
workers at Mahatma Gandhi Institute of Medical Sciences hospital, Sevagram,
India, for their participation; Padmakar Dhone, BSc, Santosh Chavhan, BSW,
Prashant Raut, BA, Vaishali Naik, BSc, Sandeep Taksande, DMLT, and Varsha
Taksande, Dip Radio, for working on this project; James Rothel, PhD, Vineet
Chadha, MD, and Hajib Madhavan, MD, for technical assistance and helpful discussions;
and Robert Jasmer, MD, Vineet Chadha, MD, Jacqueline Tulsky, MD, Dick Menzies,
MD, and Marcel Behr, MD, for reviewing the manuscript. We are also grateful
to colleagues at the US Centers for Disease Control and Prevention, for reviewing
the manuscript: Kenneth Castro, MD, Michael Iademarco, MD, Charles Wells,
MD, Kayla Laserson, ScD, Andrew Vernon, MD, and Jose Becerra, MD.