, 1997; Carbonnelle et al., 2006; Balasingham et al., 2007; Tammam et al., 2011). Structures of PilP fragments from P. aeruginosa (PDB: 2LC4) and N. meningitidis (Golovanov et al., 2006) have been solved by NMR and adopt an identical fold, but do not share the same surface properties. The N-terminus appears to lack regular secondary structure, while the C-terminus

forms a β-sandwich. The C-terminus of the PilP orthologue in E. coli T2S, GspC, has been shown to interact directly with the N0 domain of the secretin subunit (PDB: 3OSS). An additional inner membrane protein, FimV, has been shown PCI-32765 price to affect the function of T4aP and T2S in P. aeruginosa (Semmler et al., 2000; Coil & Anne, 2010; Michel et al., 2011; Wehbi et al., 2011). Mutation of fimV reduces both secretin and secretin monomer levels (Wehbi et al., 2011). No structure of FimV is yet available but the periplasmic N-terminus encodes a LysM-type PG-binding motif, while the highly acidic cytoplasmic C-terminus contains putative TPR motifs. Deletion of the LysM domain alone impairs secretin

formation, suggesting PG interactions are important for assembly. Outer membrane secretins are formed by multimerization of 12–15 molecules of a single protein into ring-like structures (Korotkov et al., 2011). The subunit in each system that forms the secretin is listed in Table 1. Secretins characterized to date can be divided into five classes: (1) self-assembling and self-membrane targeting; Carfilzomib ic50 (2) self-assembling but not self-membrane targeting; (3) self-assembling but inefficiently self-membrane targeting; (4) self-membrane targeting but not self-assembling and (5) not self-assembling and not self-membrane targeting (Fig. 2). Little correlation is readily apparent between the classes of secretins and the systems to which they belong. Class 1 secretins have only recently been identified. The single Class

1 secretin that has been characterized to date is HxcQ from the P. aeruginosa T2S. This class of secretins is unique as they are themselves lipoproteins that are directly targeted to the outer membrane by the Lol pathway, where they Bay 11-7085 auto-assemble (Viarre et al., 2009). In contrast (see below), all other secretins require additional proteins for stability, localization and/or assembly. The genomic organization of Class 1 secretins also differs from the typical Gsp-type T2S systems, where the secretin gene is usually paired with, and immediately downstream of a gspC orthologue (encoding GspC, PulC, OutC, XcpP, EtpC, or XpsN). The equivalent gene in the Hxc system, hxcP, is instead located at the opposite end of the gene cluster from the secretin gene. As GspC and the secretin in T2S are hypothesized to be responsible for co-recognition of multiple substrates (Bouley et al., 2001; Gerard-Vincent et al., 2002; Douzi et al.

39 In the second report, one passenger and one crew member were infected on an 11-hour military charter flight from the Southwestern United States to Frankfurt. One of the individuals had serogroup B disease; the serogroup in the other individual was undetermined.40 Vaccination against meningococcal disease is not required for individuals traveling into any country except for Hajj and Umrah pilgrims to Saudi Arabia.5,41 This visa requirement for Saudi Arabia has been extended to all nationals

of many countries in tropical Africa arriving by air.42 Proof of immunization is needed for all Adriamycin Hajj and Umrah visa applicants of all ages; depending on the age group and origin, other vaccinations may also be required (yellow fever, poliomyelitis, and influenza).5 Applicants must have been immunized more than 10 days and less than 3 years before entering Saudi Arabia.5 Because of this requirement for periodic vaccination, waning immunity due to immunologic hyporesponsiveness after repeat administration of meningococcal polysaccharide vaccines can be a concern.6 A study on residents of Mecca and Jeddah, Saudi Arabia, aged 10 to 29 years found that repeated administration of the AC polysaccharide vaccine resulted in immunologic hyporesponsiveness to serogroup C.6 Rapamycin order Hyporesponsiveness is not a factor with conjugate quadrivalent meningococcal vaccines,43 and they should therefore be preferred for use for instance in

Hajj pilgrims. Several national health authorities as well as the WHO have issued guidance on vaccinating travelers against meningococcal disease based on environmental factors, such as travel destination, time of year, and type of contact with the local population (Table 2). Although not all the recommendations are consistent—especially Nintedanib (BIBF 1120) in terms of the strength of the recommendation—many overlap to some degree, and all

recommend vaccination for all travelers visiting destinations with current outbreaks or epidemic situations. In its 2010 edition of International Travel and Health, the WHO lists meningococcal disease vaccination as being of selective use in travelers, along with hepatitis A vaccine, for example.44 For travelers to industrialized nations, where they may be exposed to sporadic cases of meningococcal disease, WHO cautions that risk is increased in locations where large groups of adolescents and young adults congregate, such as in schools and college dormitories, and recommends considering vaccination for college students. Vaccination should also be considered in “all travelers to countries in the sub-Saharan meningitis belt.” The risk of infection may be greater in those traveling during the dry season or those staying in the area for longer periods and living with or being in close contact with the local population.44 Vaccination is also to be considered in “all travelers … to areas with current epidemics.”44 The WHO, US, UK, and German/Swiss recommendations are very similar (Table 2).

What might be the source of face-related information for these pulvinar neurons? There are a number of possibilities that need to be considered, and, importantly, they are not mutually exclusive. The lateral pulvinar has extensive connections with the visual cortex, including the inferotemporal (IT) cortex (Shipp, 2003), where face-selective neurons have often been found clustered together, with functionally

similar neural response characteristics find more for processing of facial aspects such as gaze direction, facial expressions, and face orientation (Bruce et al., 1981; Perrett et al., 1982; Desimone et al., 1984; Pinsk et al., 2005; Tsao et al., 2006). Thus, the IT cortex is a likely source of face-related information for these pulvinar http://www.selleckchem.com/products/abt-199.html neurons. However, although face-related information in pulvinar

responses peaked at 50–100 ms in the majority of neurons, and they thus had similar response times to those of some IT cortex neurons, the response latencies of a number of these pulvinar neurons were short, the responses occurring as early as 30 ms, and the spike rate in the first 50 ms after stimulus onset provided significant information about face-like stimuli. Although it is possible that these fast pulvinar Thalidomide responses derive from the visual cortex, an alternative consideration is that these neurons receive input from an extrageniculate source of face-related information, such as the superior colliculus (SC). The pulvinar and the SC have been implicated in a fast subcortical route of face processing that provides the amygdala with input from the

SC via the pulvinar, thereby circumventing cortical processing (LeDoux, 2000). Consistent with this proposal, some of the face-related pulvinar neurons were found to be located in the medial pulvinar, the origin of pulvinar projections to the amygdala (Jones & Burton, 1976; Romanski et al., 1997). It will be interesting to explore these particular parts of the pulvinar in greater detail in future studies, and to probe aspects of face processing related to emotional valence such as fear and threat. However, others have argued against the necessity of the pulvinar providing a fast input to the amygdala, instead emphasizing a possible contribution of the pulvinar to face processing at the cortical level (Pessoa & Adolphs, 2010). Such a route may originate from the SC as well, as a disynaptic colliculo-pulvinar-cortical pathway has been shown to project to cortical areas V3 and MT (Berman & Wurtz, 2010; Lyon et al., 2010).

Sparkle (Lee & La Rue, 1992). In Trifolium repens roots, ethylene inhibits cortical cell division, a process that is indispensable for nodule primordia formation (Goodlass & Smith, 1979). To obviate some of the inhibitory effects of ethylene in nodule formation, development and function, some rhizobial strains utilize different mechanisms for lowering ethylene levels such as the production of the selleck products enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase; this enzyme is responsible

for the cleavage of ACC (the immediate precursor of ethylene in plants) to ammonia and α-ketobutyrate (Honma & Shimomura, 1978), contributing to increase the competitiveness of the strains because of advantages in the processes of nodule formation and occupancy Everolimus cell line (Ma et al., 2003b, 2004). Other rhizobial strains lower ethylene levels by producing the compound rhizobitoxine, an inhibitor of the plant enzyme ACC synthase (Sugawara et al., 2006). The prevalence of ACC deaminase genes in rhizobia has been studied primarily in Rhizobium spp. (Ma et al., 2003a; Duan et al., 2009). In these studies, many Rhizobium spp. have been found to possess an acdS gene and produce ACC deaminase under free-living conditions. For example, in a rhizobia collection of isolates from Saskatchewan (Canada), 27 Rhizobium isolates possessed an acdS gene and were able to produce

ACC deaminase, thus, showing that acdS genes are present throughout Rhizobium isolates (Duan et al., 2009). On the other hand, notwithstanding reports documenting the presence of ACC deaminase in Mesorhizobium spp., not much is known about the environmental distribution of acdS genes in this bacterial genus. The first report on acdS gene presence in Mesorhizobium was obtained following the complete sequencing of Mesorhizobium sp. MAFF303099 (Kaneko et al., 2000). Subsequently, the presence of an acdS

gene in the symbiosis island of Mesorhizobium loti R7A was also reported (Sullivan et al., 2002). However, when Mesorhizobium sp. MAFF303099 and Mesorhizobium ciceri UPM Ca-7 were tested for ACC deaminase activity and the presence of an acdS gene, no activity was detected and the acdS gene was not found in M. ciceri (Ma et al., 2003b). Recently, the genome sequences of Mesorhizobium Nintedanib datasheet opportunistum WSM2075T (Lucas et al., 2011a), Mesorhizobium australicum WSM2073T (Lucas et al., 2011b), and Mesorhizobium ciceri bv. biserrulae WSM1271 (Lucas et al., 2011c), revealed the presence of an acdS gene in these strains. In some strains of Mesorhizobium, the production of ACC deaminase has been shown to be an important mechanism to promote nodule formation. When compared to the wild-type strain, Mesorhizobium sp. MAFF303099 acdS knockout mutant has a decreased ability to form and occupy nodules, losing both its effectiveness and competitiveness (Uchiumi et al., 2004).

, 2004; Schubert et al., 2006; van Peer et al., 2010; Ohm et al., 2010). Recently, a dedicated deletion vector has been described, which

reduces screening for transformants with a gene inactivation (Ohm et al., 2010). This construct, called pDelcas, consists of two antibiotic resistance cassettes. selleck screening library The nourseothricin resistance cassette is positioned in between the flanks of the gene that is to be deleted. On the other hand, the phleomycin resistance cassette is positioned elsewhere in the construct. Consequently, phleomycin resistance is indicative of an ectopic integration of the construct. By replica plating on a medium containing phleomycin, about 70% of the transformants could be eliminated in the screening process for a strain with a gene deletion. However, 30% of the transformants still had to be screened by PCR and/or Southern hybridization. This is the reason why we decided to inactivate the ku80 gene that is part of the nonhomologous end-joining (NHEJ) pathway. The frequency of targeted gene inactivation by HR is related to the default pathway used by the organism to repair double-stranded DNA breaks (Ninomiya et al., 2004). Saccharomyces cerevisae, for instance, uses mainly HR, which is mediated by the concerted action of Rad51 and Rad52 (New

et al., 1998). This explains the high incidence of homologous integration in this organism. In most filamentous fungi, ectopic integrations are much more frequent (Fincham, 1989). Such integrations are mediated by NHEJ. NHEJ can be initiated Ruxolitinib by PARP-1, which recruits the XRCC1–DNA ligase III complex (Audebert et al., 2004). Alternatively, NHEJ results from the action of the Ku70/Ku80 heterodimer (for a review, see Weterings & Chen, 2008). This heterodimer binds to free DNA ends, and recruits and activates the DNA-dependent protein kinase catalytic

subunit. Consequently, DNA ligase IV binds to the complex formed, together with XRCC4, which results in the ligation of the DNA ends. Inactivation of ku70, ku80 or both has considerably increased targeted gene inactivation in a number of filamentous fungi (Ninomiya et al., Nintedanib (BIBF 1120) 2004; Krappmann et al., 2006; Nayak et al., 2006; Pöggeler & Kück, 2006; Takahashi et al., 2006; Choquer et al., 2008; Haarmann et al., 2008). Here, we report for the first time the inactivation of ku80 in a mushroom-forming fungus and the use of the resulting strain for the deletion of sc15 (Lugones et al., 2004) and the putative transcription factors jmj3 (containing a Jumonji DNA-binding domain) and pri2 [containing a Zn(II)2Cys6 zinc cluster DNA-binding domain]. Monokaryotic and dikaryotic strains of S. commune were grown at 25 °C in the light on minimal medium (MM; Dons et al., 1979). The monokaryotic strain H4-8 (Fowler et al., 1999) was transformed as described (van Peer et al., 2009). Twenty micrograms of vector DNA was incubated with 5 × 107 protoplasts.

Lamivudine

has been extensively used, as has tenofovir and to a lesser extent emtricitabine, for the treatment of HIV mono-infection during pregnancy, and lamivudine and telbivudine have been used in HBV mono-infected pregnant women and all have been check details found to be safe. There are limited data on adefovir use in pregnancy and it is not recommended. Where it is being used in a woman for management of HBV but who does not require HIV treatment, this should be switched to tenofovir incorporated into her HAART regimen. In the context of coinfection during pregnancy where HAART is indicated, there is unlikely to be a situation where it would be used instead of tenofovir. There is no evidence of any adverse effect on maternal health if women become pregnant while taking tenofovir, lamivudine or emtricitabine: these drugs are recommended as NRTI choices in national [10] and international guidelines [2]. 6.1.6 In all HAV non-immune HBV coinfected women, HAV vaccine is recommended after the first trimester as per the normal schedule (0 and 6–12 months) unless the CD4

cell count is <300 cells/μL, when an additional dose may be indicated. Grading: 1D Immunization for HAV uses inactivated vaccines. Data for HAV vaccine in pregnancy are limited. Nevertheless, several guidelines indicate that pregnancy is not a contraindication for HAV immunization, including in HBV coinfected pregnant women [[11],[12]]. Montelukast Sodium For HAV vaccines, patients with higher CD4 cell counts and on HAART generally show improved responses to selleck vaccination. HIV-positive persons with CD4 cell counts <300 cells/μL should receive three doses of HAV vaccine over 6–12 months instead of the standard two. 6.1.7 Tenofovir and emtricitabine should form the backbone of an ART regimen in naïve patients with wild-type HIV/HBV infection and no contraindication to either drug (Grading: 1B). 6.1.8 If tenofovir is not

currently part of HAART it should be added. Grading: 1B 6.1.9 Lamivudine/emtricitabine may be omitted from the ARV regimen and tenofovir given as the sole anti-HBV agent if there is clinical or genotypic evidence of lamivudine/emtricitabine resistant HBV. Grading: 1C 6.1.10 Lamivudine or emtricitabine should not be used as the only active drug against HBV in HAART because of the likelihood of emergent HBV resistance to these agents. Grading: 1B 6.1.11 Emtricitabine has potential antiviral benefits over lamivudine, is coformulated with tenofovir, and appears to be equally safe during pregnancy and hence is the preferred option to be given with tenofovir in coinfection. Grading: 2D All HBV/HIV coinfected women should receive HAART containing tenofovir with emtricitabine or lamivudine treatment during pregnancy, unless contraindicated.

To combine these two separate experimental data, event frequencies should be normalised by the unit length of the axon (axonal short-pause rates, axonal appearance and disappearance rates; see ‘Materials and methods’; Fig. 8). The axonal appearance and disappearance rates were measured from the same experimental

data shown in Fig. 3 (Fig. 1C). The short-pause rate of individual mitochondria was suppressed by TTX treatment at 3 weeks (Fig. 5B). However, the axonal short-pause rate was not changed by TTX treatment because the number of mobile mitochondria was increased by TTX treatment (Figs 3I and 8). By using these normalised rates, we could calculate the stabilisation rates at different conditions ([SPSS]; Fig. 8). The stabilisation rate Sirolimus near synapses ([SPSS]synaptic) declined significantly from 2 to 3 weeks (1.01 vs. 0.53%) and was modulated by TTX treatment. Because stabilisation rates away from synapses ([SPSS]non-synaptic) were less affected by culture periods and TTX treatment, regulation of the stabilisation rate near synapses is likely SP600125 molecular weight to be the parameter that is important for the control of mitochondrial replacement along the axon. Although the axonal appearance rate of

mitochondria near synapses ([MSS]synaptic) was more than twofold higher at 2 weeks, this increase was counterbalanced by the comparable rate of disappearance ([SSM]synaptic). It is likely that there exists a mechanism that keeps the balance between [MSS] and [SSM], as these rates were maintained in parallel in all experimental conditions (Fig. 8). This regulation may be important to keep the density of both synaptic and non-synaptic mitochondria constant with time. We report here the dynamic properties of axonal mitochondria using live-cell imaging with multiple sampling frequencies ranging from seconds to days. High-frequency image sampling is necessary to trace the accurate positions of mobile mitochondria, transported by motor proteins with their velocity of 0.1–1.4 μm/s (De Vos & Sheetz, 2007; MacAskill & Kittler, 2010).

In turn, the probability of transitions between stationary and mobile states is low (a few events per hour within an image area; Fig. 8) and time-lapse imaging with longer durations is required. Here we performed time-lapse imaging with high (intervals of 3 s), intermediate (intervals of 30 min) selleck compound and low (intervals of 1 day) frequencies. Our results demonstrated that mitochondrial dynamics on multiple time scales differ between developmental stages and are regulated by neuronal activity and proximity to synaptic sites. To understand the dynamics of axonal mitochondrial distribution, mitochondrial properties in mobile and stationary states, and the transition process between them should be examined (Fig. 1). Our analyses revealed that the properties of stationary mitochondria are highly regulated by neuronal maturation and activity.

Currently, instituting a second pharmacy check of PTTAs is not warranted. 1. Dodds LJ.

Pharmacist contributions to ensuring safe and accurate transfer of written medicines-related discharge information: lessons from a collaborative audit 5FU and service evaluation involving 45 hospitals in England. Eur J Hosp Pharm Published Online First: 10 February 2014. doi:10.1136/ejhpharm-2013-000418 K. Medlinskiene Hull and East Yorkshire NHS Hospitals, Hull, UK HDS is the main communication tool between hospital and general practitioners. Evaluate turnaround time for HDS and to what extent pharmacist input was required. The average turnaround time for HDS in the pharmacy was 2 h 22 min and 75% of HDS required pharmacist input. The hospital discharge summary

learn more (HDS) is the main method of communicating patient’s diagnostic findings, hospital management, and arrangements for post-discharge follow up to general practitioners. HDS are additionally checked by hospital pharmacists if discharge medication supply is required. It is not unusual to receive complaints from patients about long waiting times for discharge medication. The study aimed to evaluate average time of a HDS journey and extent to which pharmacist input was required. The data collection was performed during one week in November 2013 at one of three acute NHS Trust sites. All HDS received in the pharmacy had forms attached for time recordings (time a HDS was created, reached the pharmacy,

turnaround time in the pharmacy). Data from HDS with completed time recordings was retrospectively analysed with Microsoft Excel to evaluate if pharmacist input was required. Any interventions, contributions and adjustments to HDS e.g. dose changes, additional instructions, completion of stopped medication box, completion of allergy status, were classed as pharmacist input. Ethical approval was not required. A total of 196 HDS had completed forms which represented 62% (314) of all HDS received that week Sitaxentan by the pharmacy. The average time for one HDS to reach the pharmacy once it had been created was 1 h 4 min. Only 5% (10) HDS were in the pharmacy 24 h prior discharge as per trust policy.1 The average turnaround time for a HDS was 2 h 22 min, which was considerably lower on the weekend (1 h 18 min). Each HDS was collected or delivered to the ward on average within 33 min. The overall average time of HDS journey was 3 h 59 min. The majority of HDS, 75% (147), required pharmacist input. Pharmacist input was achieved by using information on inpatient drug cards, contacting ward (nurse or doctor), or both (Table 1). Table 1 Sources used for pharmacists input Drug card 70% (103) Contacting ward (doctor or nurse) 2% (3) Both 28% (44) HDS are mostly written by junior doctors and errors are often associated with this junior status.

Although DY380 works well for most experiments, it, however, must be propagated at 30 °C and a precise and homogenous water bath is required for the 15-min heat induction of λ Red genes. The third is the integrative form system. Representative strains in the system are KM22 (Murphy, 1998) and YZ2000 (Zhang et al., 2000). KM22 was obtained by replacing the cellular RecBCD genes of E. coli AB1157 with exo and bet under the lac promoter control. YZ2000 was generated by deleting the restriction/modification systems and the endogenous Staurosporine chemical structure lac operon of sbcA strain JC8679. YZ2000 functions through the recE and recT genes originating

from the E. coli chromosomal lambdoid Rac prophage, and recET shows the same yet less efficient enzymatic functions as their counterparts exo and bet (Muyrers et al., 2000); still, YZ2000 may degrade the incoming DNA for the lack of gam. As λ Red recombineering is now often used to modify large constructs such as BAC (bacterial artificial buy Y-27632 chromosome), YZ2000 and KM22 may be inferior to the E. coli DH10B-based host strains that are used for large construct propagation. Each recombineering system has its advantage. The

advantage of the plasmid-based recombineering system is that the plasmid can be transformed into any E. coli host strain as long as it can coexist with the targeting DNA, while the advantage of phage-based and integrative form systems is that the recombineering function does not rely on plasmids, which means that no plasmid introduction or

plasmid elimination is needed in the transformation procedure. Among the three recombineering systems, the integrative form is the least often used one. To make the best use of the integrative form system, in this study, we engineered a new recombineering strain LS-GR by integrating the functional recombineering elements, including the λ Red genes, recA, araC and aacC1 (gentamicin resistance gene), into the E. coli DH10B chromosome. recA, when incorporated as the transient expression of recA into the plasmid-based recombineering system, has been demonstrated to improve the recombination efficiency significantly (Wang et al., 2006) and Ceramide glucosyltransferase the recA mutant strain led to 68-fold less recombination efficiency (Murphy, 1998). The recombineering function of LS-GR was characterized through pACYC184 and pECBAC1 modifications. The same modifications with pKD46 and pSC101-BAD-gbaA as recombineering function suppliers were performed in parallel to evaluate the recombination efficiency of LS-GR. Plasmid pBAD322G (Cronan, 2006) containing aacC1 was obtained from John Cronan. pACYC184 is a p15A replicon origin, medium copy number (10–15 copies) vector. Single copy number BAC vector pECBAC1 (Frijters et al., 1997) was obtained from Richard Michelmore. Escherichia coli BW25141/pKD4 and E. coli BW25113/pKD46 (Datsenko & Wanner, 2000) were obtained from Barry Wanner through E. coli Genetic Stock Center, Yale University.

0; SPSS Inc., Chicago, IL). The differences in the species-specificity and the limit of detection between the different bacterial samples were evaluated using Student’s t-tests. All the 33 isolates of S. pyogenes and the test strains were amplified using H2 primer. The primer produced RAPD patterns consisting of two to eight distinct DNA fragments,

generally ranging from approximately 400 to 2000 bp. A reference strain [S. pyogenes (GAS SF370)] was used in the analysis and produced two prominent bands of approximately 400 and 1400 bp (Fig. 1). Similar patterns were observed in all 33 isolates of the present study. Eight different RAPD profiles (designated A–H) were found among buy MS-275 the 33 S. pyogenes isolates. RAPD profile A was predominant and observed in 14 isolates (represented by lanes 1, 2 and 6) (Fig. 1) followed by F and G, with 10 (lane 8) and three isolates (lane 9), respectively. Profile B (lane 3), C (lane 4), D (lane 5), E (lane 10) and H (lane 11) were represented by an isolate each. The genomic fingerprints produced by H2 primer gave rise to reliable and reproducible polymorphic

fragments of 400 and 1400 bp in length. R428 in vivo In the development of a species-specific marker for S. pyogenes, the 419-bp monomorphic band (hereafter referred as MB) was chosen, and then cloned, sequenced and deposited in the EMBL/GenBank/DDBJ databases (EU660382). This sequence partially codes for an enzyme 3-keto acyl reductase. The presence of MB was confirmed through RAPD with the test strains; none of these strains possessed this fragment (Fig. 2). In particular, the MB was not present in other species of the same genus (GBS, GCS and GGS). The MB was highly specific to S. pyogenes, which showed the closest match of 98% similarity. The SCAR primers were designed within a region of the MB. The SCAR primers were named on the basis of the expected length of amplified product. The annealing temperature and the MgCl2 concentration were optimized at 60 °C and 1.5 mM, respectively, to adjust for the stringency of PCR conditions, thus minimizing the possibility of nonspecific hybridization with nontarget

DNA. The primer pair was evaluated against the test strains and different Streptococcus species. The 212F/212R primer pair gave rise to a single, strain-specific amplification product, which Megestrol Acetate was used for subsequent analysis. The specificity of SCAR primers 212F/212R was evaluated against DNA extracted from the clinical isolates of S. pyogenes and non-GAS test strains. The results indicated that the primers were highly specific for amplifying genomic DNA from all 33 S. pyogenes isolates. The efficiency of the primers when analysed against the non-GAS test strains showed amplification only in the positive control SF370. The sensitivity of the SCAR primers was tested by qualitative PCR. The sensitivity in nanograms of target DNA per PCR was evaluated by means of artificial mixtures prepared by adding known aliquots (102–10−3 ng−1 PCR) of genomic DNA of S. pyogenes.