Comparative Mycobacterium tuberculosis Spoligotype Distribution in Mexico

Transcription

1 JCM Accepts, published online ahead of print on 21 May 2014 J. Clin. Microbiol. doi: /jcm Copyright 2014, American Society for Microbiology. All Rights Reserved Comparative Mycobacterium tuberculosis Spoligotype Distribution in Mexico LUCIO VERA-CABRERA 1 *, JESSICA RAMOS-ALVAREZ 3, CARMEN A. MOLINA- TORRES 1, LYDIA GUADALUPE RIVERA-MORALES 3, ADRIÁN RENDÓN 3, FRANCISCO QUIÑONES-FALCONI 4, JORGE OCAMPO-CANDIANI 1. 1 Servicio de Dermatología, Hospital Universitario, UANL, Monterrey, Mexico; 2 Centro de Investigación, Prevención y Tratamiento de Infecciones Respiratorias, Hospital Universitario, UANL, Monterrey, México; 3 Departamento de Inmunología, Facultad de Ciencias Biológicas, UANL, Monterrey, México; 4 Servicio de Microbiología Clínica, Instituto Nacional de Enfermedades Respiratorias, Secretaría de Salud, México. Keywords: Tuberculosis, Spoligotypes, México *Corresponding author Mailing address: Dr. Lucio Vera-Cabrera, Servicio de Dermatologia, Hospital Universitario, Madero y Gonzalitos, Colonia Mitras Centro, C.P , Monterrey, N.L., México. Phone: (81) Fax: (81) drluciovera@yahoo.com.

2 ABSTRACT In the present work, we studied the genetic diversity of Mycobacterium tuberculosis clinical isolates according to the gender, age, and geographic location within Mexico. We did not observe any statistically significant difference considering age and gender. We found that SIT53 is more frequent in the northern states and that SIT119 predominates in central Mexico

3 In a previous work, we described the spoligotype distribution in the Nuevo Leon state and reported that SIT53 and SIT119 were the most common spoligotypes to be isolated (representing approximately 39% of the total sample) (1). We also reported the tendencies of SIT53 to be isolated more frequently in elderly patients and of SIT119 to be isolated more frequently in younger patients (<20 years old). However, our sample size was too small to definitively support this conclusion. In the present work, we used a larger population sample to determine whether this difference is genuine and compared the distribution of spoligotypes in Nuevo Leon against those of the oldest and most densely populated region of our country, Mexico City, which has a population of approximately 30 million people. The M. tuberculosis isolates (n=411) were obtained from the laboratories at either the Laboratorio Estatal de Salud Pública or the tuberculosis clinic at the Hospital Universitario, José E. Gonzalez U.A.N.L, in Monterrey, Mexico, from 2001 to Because in a previous report we observed a relationship between the distribution of spoligotypes and the patients gender and age, we included 210 new samples from Nuevo León with the following characteristics: 116 cases were from patients aged 0-20 years old, and 117 cases were from patients aged over 50 years. From the Instituto Nacional de Enfermedades Respiratorias (INER) in Mexico City, we obtained 201 patients from several of Mexico s states, including Distrito Federal (n=126), Chiapas (n=10), Oaxaca (n=11), Veracruz (n=24), San Luis Potosí (n=1), Jalisco (n=1), and Michoacán (n=8). Isolates included the open population and were grown on Lowenstein-Jensen medium. The DNA isolation was performed using the method of Van Embden et al (2). Spoligotyping was accomplished using a standard technique, as described previously (3).

4 The spoligotypes were entered into the SITVIT2 database (Pasteur Institute of Guadeloupe) in a binary format. This database is an updated version of the previously released SpolDB4 database (available at To establish differences in the spoligotype distribution among patients of different ages and genders, 251 samples corresponding to individuals who were either <30 or >60 years of age were analyzed. We observed a difference of 33% for SIT119 over SIT53 in the male patients, although this difference was not statistically significant. In the groups divided by age, we observed a 33% increase in the number of SIT119 spoligotypes in the younger patients; however, no significant differences were observed (Table 1). Finally, we identified no significant correlation between age/gender for any of the other spoligotype groups. A total of 411 tuberculosis isolates were collected from 12 Mexican states. Most of these samples were collected from Nuevo Léon state and México City. We observed 91 spoligotype patterns, which were classified into 13 clades and 32 subclades (Table 1S, supplemental material). The most frequently identified SITs were 119, 53, 50, and 42 in the families X1, T1, H3, and LAM9, respectively (Fig 1). When comparing Nuevo Leon and Mexico City, we found that SIT119 was significantly more predominant in the latter, with SIT53 as the second most common isolate. Interestingly, SIT119 was only reported in Mexico City, the State of Mexico (adjacent to Mexico City), and the state of Nuevo Leon, but not in any other regions of Mexico (4,5,6,7,8,9,10)(Fig 2). By contrast, SIT53 has been found to be abundant throughout the republic. When compared to other regions of the American continent (Table 2), SIT53 has been reported to be dominant only in a

5 single study in Sao Paulo, Brazil, but scarce in the rest of South America (11, 12, 13, 14,15) We also identified 7 isolates of SIT1 (the Beijing family) (1.7%) and 5 isolates of M. bovis (1.2%). in order to classify the lineage of our seven Beijing strains, we analyzed the presence of genetic deletions by PCR (16). When subtyped, all of the strains showed the following deletions: RD105, RD149, RD 152, RD207, and RD181. Notably, the RD150 deletion was absent in four strains, and only one strain exhibited the RD142 deletion. These results indicate that the isolated Beijing strains do not share the same origin and that they correspond to different sublineages In this study, we observed that the most predominant TB subtype is SIT53; this SIT has also been found to be the most common in the nearby region of San Luis Potosi (600 km from Monterrey) (7), and Guadalajara (near the Pacific side of the country) (8) (See map in Fig 2). In Mexico City, SIT53 has been reported to be the predominant subtype in patients with immunodeficiency (5) and MDR isolates (4). In HIV patients and pediatric patients (5,6), the LAM9 family spoligotype SIT42 predominates, with SIT53 as the next most common spoligotype. Beyond Mexico City, SIT53 is quite predominant throughout the country; depending on the region, there is a second dominant subtype, either SIT42 or SIT119. SIT53 is also commonly found in Spain (Madrid), the country responsible for colonizing most of America during the 1500s (16). When comparing with neighboring United States, we observe that in Houston, the largest city close to Monterrey, the predominant SIT belongs to the Beijing family (22.8% vs. 3.2% of SIT53) (18). The diversity of

6 the dominant spoligotypes in America might reflect their original dissemination through the Europeans who came to the continent. ST119 is the second most common spoligotype in Monterrey, and it has been observed in Mexico City, but not in the other states of the republic. Monterrey City was founded in 1596 by Jewish refugees from the Spanish Inquisition (19). The other group that comprises a significant portion of this city s population is the Amerindian Tlaxcaltecan group, which was given the land by the Spaniards in exchange for, help conquering the Aztec empire. Before these two groups of settlers, the population was quite low in this region and was constituted mainly of nomadic people. To date, however, Monterey is the second/third largest city in Mexico, which attracts people from other states. It possibly that it was brought to Monterrey by a group that did not migrate to the rest of the republic. When assessing the relationship between spoligotype and patient age, we observed that SIT119 is more common in young males than in persons aged >60 years. However, this difference was not statistically significant. Most cases in elderly people are endogen or latent tuberculosis cases that were acquired a long time ago, while cases in young people instead represent recent transmission or disease acquisition. It is thus possible that SIT119 is emerging in the most affected group (males) and that it will constitute the most dominant spoligotype in the future. It will be interesting to analyze the spoligotype distribution in Monterrey in 10 years and to determine the further evolution of these subtypes in the years to come

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9 Gomes HM, Elias AR, Oelemann MA, Pereira MA, Montes FF, Marsico AG, Kritski AL, Filho Ldos A, Caldas PC, Possuelo LG, Cafrune P, Rossetti ML, Lucena N, Saad MH, Cavalcanti HR, Leite CQ, de Brito RC, Lopes ML, Lima K, Souza M, Trindade Rde C, Zozio T, Sola C, Rastogi N, Suffys PN Spoligotypes of Mycobacterium tuberculosis complex isolates from patients residents of 11 states of Brazil. Infect. Genet. Evol. 12: Sheen P, Couvin D, Grandjean L, Zimic M, Dominguez M, Luna G, Gilman RH, Rastogi N, Moore DA Genetic diversity of Mycobacterium tuberculosis in Peru and exploration of phylogenetic associations with drug resistance. PLoS One. 8:e Abadía E, Sequera M, Ortega D, Méndez MV, Escalona A, Da Mata O, Izarra E, Rojas Y, Jaspe R, Motiwala AS, Alland D, de Waard J, Takiff HE Mycobacterium tuberculosis ecology in Venezuela: epidemiologic correlates of common spoligotypes and a large clonal cluster defined by MIRU-VNTR-24. BMC. Infect. Dis. 9: Rosales S, Pineda-García L, Ghebremichael S, Rastogi N, Hoffner SE Molecular diversity of Mycobacterium tuberculosis isolates from patients with tuberculosis in Honduras. BMC Microbiol. 10: Cerezo I, Jiménez Y, Hernandez J, Zozio T, Murcia MI, Rastogi N A first insight on the population structure of Mycobacterium tuberculosis complex as studied by spoligotyping and MIRU-VNTRs in Bogotá, Colombia. Infect. Genet. Evol. 12:657-63

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11 ACKNOWLEDGMENTS This work was supported by the Consejo Nacional de Ciencia y Tecnologia (CONACYT) grant No , and PAICYT SA This work was supported by the Apoyo a la Incorporación de Nuevos PTC del Programa de Mejoramiento al Profesorado grant PROMEP/103-5/11/4330, from Secretaría de Educación Pública

12 FIGURE FOOTNOTES Fig. 1. Frequency of SITs found in 12 states of México. The pie chart includes all the isolates analyzed in this study (n=411); in the right panel, we show the distribution according to the geographical origin of the patients, but show only the most predominant spoligotypes: SIT53, SIT119 and SIT50.

13 13 SIT8 SIT19 SIT42 SIT53 SIT119 n>100 n>50 n<50 Monterrey, Nuevo León MDR strains Pediatric pa ents San Luis Potosí México City HIV pa ents Inmunodeficient pa ents Guadalajara, Jalisco México state Veracruz Acapulco, Guerrero Fig. 2. Frequency of SITs reported in México. Here we observe the distribution of the 5 most commonly reported SIT s in previous studies in the Mexican republic

14 TABLES

15 Table 1. Distribution of spoligotypes according to patient gender and age. The statistical analysis with Students s T test did not show a statistically significant difference in any case Most frequent SIT M F > 50 years old < 30 years old SIT SIT SIT SIT

16 16 Table 2.- Comparative table between spoligotypes found in several American countries and from Spain. 433

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