Dennis Nurjadi

Import and spread of Panton-Valentine leukocidin-positive Staphylococcus aureus through nasal carriage and skin infections in travelers returning from the tropics and subtropics.

BACKGROUND  Antibiotic-resistant Staphylococcus aureus is a globally emerging pathogen. Exchangeable virulence factors, such as Panton-Valentine leukocidin (PVL), have been proposed to drive this epidemic. We investigated whether skin infections and nasal colonization in travelers contribute to the global spread of such strains. METHODS  We conducted a case-control study of 38 returnees from the tropics and subtropics with S. aureus-positive skin and soft tissue infections (SSTIs) and 124 control patients with other travel-associated disorders. We collected information on travel characteristics, clinical outcomes of SSTIs, antibiotic sensitivity patterns, and genotypes of S. aureus strains isolated from skin lesions and the nares. RESULTS  S. aureus-positive SSTIs were associated with travel duration and purpose and were most common in returnees from Africa (odds ratio, 4.2; P = .005). PVL-positive (PVL(+)) S. aureus was frequent in the lesional and nasal isolates from travelers with SSTIs but could not be found in the nares of the control patients. The presence of PVL in S. aureus in travelers was associated with complicated disease, reduced antibiotic susceptibility, and secondary spread. The genotypes of PVL(+) S. aureus in returnees were reported to be endemic to the visited destination but rarely observed in Europe. CONCLUSIONS  Geographic variation in the risk of SSTIs in travelers supports a globally heterogeneous distribution of virulent S. aureus. Complicated SSTIs in returnees from nontemperate climates are associated with PVL(+) S. aureus and promote the emergence and spread of virulent and antibiotic-resistant strains. We propose a network for the surveillance of imported S. aureus (www.staphtrav.eu).

Impaired β-Defensin Expression in Human Skin Links DEFB1 Promoter Polymorphisms With Persistent Staphylococcus aureus Nasal Carriage

BACKGROUND Genetically determined variation in the expression of innate defense molecules may explain differences in the propensity to be colonized with Staphylococcus aureus. METHODS We determined S. aureus nasal carriage in 603 volunteers; analyzed polymorphisms in the DEFB1 promoter at positions -52 G>A (rs1799946), -44 C>G (rs1800972), and -20 G>A (rs11362); and measured the content of human β-defensin 1 (hBD-1) and hBD-3 messenger RNA (mRNA) in 192 samples of healthy and experimentally wounded human skin. RESULTS Compared with GGG at the positions -52/-44/-20, the ACG haplotype was more common among persistent S. aureus nasal carriers (odds ratio, 1.93; 95% confidence interval [CI], 1.2-3.1; P = .006) and was associated with reduced expression of hBD-1 (GGG>ACG>GCA; P < .001) and hBD-3 (GGG>GCA>ACG; P = .04) in skin when measured 72 hours after wounding. Furthermore, a 50% decrease in hBD-1 and hBD-3 mRNA expression in wounded skin increased the odds of persistent carriage by 1.45 (95% CI, .93-2.26; P = .1) and 1.48 (95% CI, 1.01-2.17; P = .04), respectively. Adjustment for known risk factors of persistent S. aureus carriage did not substantially change the associations of both DEFB1 haplotypes and β-defensin expression with S. aureus colonization. CONCLUSIONS DEFB1 polymorphisms may promote persistent S. aureus colonization by altering β-defensin expression in keratinocytes of human skin.

Persistent Nasal Carriage of Staphylococcus aureus Is Associated with Deficient Induction of Human β-Defensin 3 after Sterile Wounding of Healthy Skin In Vivo

ABSTRACT Persistent nasal carriage of Staphylococcus aureus is the primary reservoir for this pathogen and a risk factor for infection. The nares of 12 to 30% of healthy individuals are persistently colonized with staphylococci. Elucidating the yet enigmatic determinants of this phenomenon is of major public health interest. We hypothesized that differences in the levels of antimicrobial peptides (AMPs) that are found in human skin and have pronounced antistaphylococcal activity may contribute to this phenomenon. We compared constitutive and induced mRNA levels of RNase 7 and human β-defensin 3 (HBD-3) in healthy and experimentally wounded gluteal skin of 60 volunteers after ascertaining their carrier status through repeated nasal cultures. We found that levels of HBD-3 expression in skin of persistent nasal carriers of S. aureus were lower: induced levels in carriers were 63% (95% confidence interval, 43 to 94%; P = 0.02) and constitutive levels were 76% (95% confidence interval, 52 to 110%; P = 0.14) of those found in noncarriers. No such associations were present for RNase 7. In conjunction with existing knowledge, these findings suggest that healthy individuals with deficient HBD-3 expression in keratinocytes are more prone to persistent nasal colonization with S. aureus.

Skin and soft tissue infections in intercontinental travellers and the import of multi-resistant Staphylococcus aureus to Europe

Staphylococcus aureus is emerging globally. Treatment of infections is complicated by increasing antibiotic resistance. We collected clinical data and swabs of returnees with skin and soft tissue infections (SSTI) at 13 travel-clinics in Europe (www.staphtrav.eu). Sixty-two percent (196/318) SSTI patients had S. aureus-positive lesions, of which almost two-thirds (122/196) were Panton-Valentine leukocidin (PVL) positive. PVL was associated with disease severity, including hospitalization for SSTI (OR 5.2, 95% CI 1.5-18.2). In returnees with SSTI, longer travel and more intense population contact were risk factors for nasal colonization with PVL-positive S. aureus. Imported S. aureus frequently proved resistant to trimethoprim-sulfamethoxazole (21%), erythromycin (21%), tetracycline (20%), ciprofloxacin (13%), methicillin (12%) and clindamycin (8%). Place of exposure was significantly (p < 0.05) associated with predominant resistance phenotypes and spa genotypes: Latin America (methicillin; t008/CC24/304), Africa (tetracycline, trimethoprim-sulfamethoxazole; t084/CC84, t314/singleton, t355/CC355), South Asia (trimethoprim-sulfamethoxazole, ciprofloxacin; t021/CC21/318), South-East Asia (clindamycin; t159/CC272). USA300-like isolates accounted for 30% of all methicillin-resistant S. aureus imported to Europe and were predominantly (71%) acquired in Latin America. Multi-resistance to non-β-lactams were present in 24% of imports and associated with travel to South Asia (ORcrude 5.3, 95% CI 2.4-11.8), even after adjusting for confounding by genotype (ORadjusted 3.8, 95% 1.5-9.5). Choosing randomly from compounds recommended for the empiric treatment of severe S. aureus SSTI, 15% of cases would have received ineffective antimicrobial therapy. These findings call for the development of regionally stratified guidance on the antibiotic management of severe imported S. aureus disease and put the infected and colonized traveller at the centre of interventions against the global spread of multi-resistant S. aureus.

Emergence of trimethoprim resistance gene dfrG in Staphylococcus aureus causing human infection and colonization in sub-Saharan Africa and its import to Europe

OBJECTIVES Co-trimoxazole (trimethoprim/sulfamethoxazole) is clinically valuable in treating skin and soft tissue infections (SSTIs) caused by community-associated methicillin-resistant Staphylococcus aureus (MRSA). The genetic basis of emerging trimethoprim/sulfamethoxazole resistance in S. aureus from Africa is unknown. Such knowledge is essential to anticipate its further spread. We investigated the molecular epidemiology of trimethoprim resistance in S. aureus collected in and imported from Africa. METHODS Five hundred and ninety-eight human S. aureus isolates collected at five locations across sub-Saharan Africa [Gabon, Namibia, Nigeria (two) and Tanzania] and 47 isolates from travellers treated at six clinics in Europe because of SSTIs on return from Africa were tested for susceptibility to trimethoprim, sulfamethoxazole and trimethoprim/sulfamethoxazole, screened for genes mediating trimethoprim resistance in staphylococci [dfrA (dfrS1), dfrB, dfrG and dfrK] and assigned to spa genotypes and clonal complexes. RESULTS In 313 clinical and 285 colonizing S. aureus from Africa, 54% of isolates were resistant to trimethoprim, 21% to sulfamethoxazole and 19% to trimethoprim/sulfamethoxazole. We found that 94% of trimethoprim resistance was mediated by the dfrG gene. Of the 47 S. aureus isolates from travellers with SSTIs, 27 (57%) were trimethoprim resistant and carried dfrG. Markers of trimethoprim resistance other than dfrG were rare. The presence of dfrG genes in S. aureus was neither geographically nor clonally restricted. CONCLUSIONS dfrG, previously perceived to be an uncommon cause of trimethoprim resistance in human S. aureus, is widespread in Africa and abundant in imported S. aureus from ill returning travellers. These findings may foreshadow the loss of trimethoprim/sulfamethoxazole for the empirical treatment of SSTIs caused by community-associated MRSA.

Hormonal Contraceptive Use and Persistent Staphylococcus aureus Nasal Carriage

BACKGROUND Human nares colonized with Staphylococcus aureus are the most important reservoir for this pathogen. We studied the influence of sex and hormonal contraceptive use on persistent S. aureus nasal carriage. METHODS We conducted a cohort study in healthy volunteers and determined carriage status at baseline and again at follow-up by using the results of 2 swab samples at each time point. We applied logistic regression to analyze associations of interest. RESULTS At baseline, 266 of 1180 volunteers (22.5%) were classified as persistent nasal carriers. Compared with women not using hormonal contraceptives, women taking reproductive hormones (odds ratio [OR]. 1.88; 95% confidence interval [CI], 1.29-2.75; P = .001) and men (OR., 1.57; 95% CI, 1.08-2.28; P = .02) were more likely to be persistent carriers. These associations remained stable after adjusting for known risk factors of nasal carriage. Women taking hormonal contraceptives and being persistent carriers at baseline were more likely to remain carriers after a median follow-up time of 70 days than women not using such medication (OR, 3.25; 95% CI, 1.44-7.34; P = .005). No patterns of association could be observed between persistent carriage among women and type of progestin or dose of estrogen used. Assuming causality and using estimates from multivariable logistic regression, we approximated that 20% (95% CI, 2.4%-34.9%) of persistent nasal carriage among women represented by our sample is attributable to hormonal contraception (population-attributable fraction). CONCLUSIONS The widespread use of hormonal contraception may substantially increase the human S. aureus reservoir with potential impact on S. aureus infection and transmission.

Staphylococcus aureus throat carriage is associated with ABO-/secretor status.

OBJECTIVES In 30% of carriers, Staphylococcus aureus colonization affects exclusively the pharynx and occurs independently from its presence in the nares. This additional reservoir has implications for S. aureus transmission, infection, and decolonization. Host factors promoting colonization of the throat, however, are unknown. METHODS We determined pharyngeal and persistent nasal carriage of S. aureus, ABO histo-blood group and ABH secretor status phenotypes in 227 individuals. RESULTS Compared to group A/non-secretors, group O/non-secretor individuals were at increased risk of carrying S. aureus in their throat (OR 6.50, 95% confidence interval 1.28-33.03, P = 0.02) and group O/secretor individuals were protected (OR 0.24, 0.07-0.77, P = 0.02). Both associations became moderately stronger after adjusting for persistent S. aureus nasal carriage, which was found to be a risk factor for pharyngeal colonization in the univariable analysis (OR 2.41, 1.35-4.33, p = 0.003). Most simultaneous carriers (72%) had identical S. aureus genotypes in their nose and throat. CONCLUSIONS These findings are consistent with in vitro studies that proposed a role of histo-blood group antigens as ligands for S. aureus and support their contribution to the observed population variation in nasopharyngeal S. aureus colonization. Based on their tissue specific expression histo-blood group antigens appear to modulate individual S. aureus colonization patterns.

Clonal expansion accounts for an excess of antimicrobial resistance in Staphylococcus aureus colonising HIV-positive individuals in Lagos, Nigeria

Nasal colonisation with Staphylococcus aureus is a risk factor for invasive infection in human immunodeficiency virus (HIV)-positive individuals. This study aimed to characterise colonising S. aureus from regions with a high HIV prevalence. Single nasal swabs were taken from a total of 374 HIV-positive and 370 healthy individuals. Overall, 202 S. aureus carriers were detected. Compared with healthy individuals, HIV-positive subjects were more likely to be S. aureus nasal carriers (33% vs. 21%; P=0.0001). Isolates from HIV-positive individuals were more often resistant to meticillin (16% vs. 8%; P=0.13), chloramphenicol (47% vs. 16%; P<0.0001), sulfamethoxazole/trimethoprim (SXT) (90% vs. 55%; P<0.0001) and ciprofloxacin (18% vs. 0%; P<0.0001). Strains belonging to the spa clonal complexes 3772/ST25 and 064/ST8 were significantly more often isolated from HIV-positive individuals and exhibited greater resistance to ciprofloxacin, SXT and chloramphenicol (spa-CC 3772) or to meticillin (spa-CC 064), respectively. Panton-Valentine leukocidin gene content was high overall and was equally distributed between isolates from HIV-positive and healthy individuals (33% vs. 30%). Genotypic characteristics of colonising isolates were similar to those reported to cause invasive infection in Nigeria. The HIV pandemic contributes to the evolution of antimicrobial resistance in S. aureus. Measures to contain antimicrobial resistance of S. aureus in Nigeria must target risk groups such as HIV-positive individuals.

Reliable and rapid characterization of functional FCN2 gene variants reveals diverse geographical patterns

BackgroundFicolin-2 coded by FCN2 gene is a soluble serum protein and an innate immune recognition element of the complement system. FCN2 gene polymorphisms reveal distinct geographical patterns and are documented to alter serum ficolin levels and modulate disease susceptibility.MethodsWe employed a real-time PCR based on Fluorescence Resonance Energy Transfer (FRET) method to genotype four functional SNPs including -986 G > A (#rs3124952), -602 G > A (#rs3124953), -4A > G (#rs17514136) and +6424 G > T (#rs7851696) in the ficolin-2 (FCN2) gene. We characterized the FCN2 variants in individuals representing Brazilian (n = 176), Nigerian (n = 180), Vietnamese (n = 172) and European Caucasian ethnicity (n = 165).ResultsWe observed that the genotype distribution of three functional SNP variants (−986 G > A, -602 G > A and -4A > G) differ significantly between the populations investigated (p < 0.0001). The SNP variants were highly linked to each other and revealed significant population patterns. Also the distribution of haplotypes revealed distinct geographical patterns (p < 0.0001).ConclusionsThe observed distribution of the FCN2 functional SNP variants may likely contribute to altered serum ficolin levels and this may depend on the different disease settings in world populations. To conclude, the use of FRET based real-time PCR especially for FCN2 gene will benefit a larger scientific community who extensively depend on rapid, reliable method for FCN2 genotyping.