SUMMARYMecA, a structural gene located on the chromosome of Staphylococcus aureus, characterizes methicillin-resistant S. aureus (MRSA), and femA and femB(fem) genes encode proteins which influence the level of methicillin resistance of S. aureus. In order to examine effectiveness of detecting mecA and fem genes in identification of MRSA, the presence of these genes in 237 clinically isolated strains of staphylococci was investigated by polymerase chain reaction (PCR). An amplified mecA DNA fragment of 533 base pairs (bp) was detected in 100 % of oxacillin-resistant S. aureus, in 16'7 % of oxacillin-sensitive S. aureus, in 81-5 % of S. epidermidis, and in 58'3 % of other coagulase-negative staphylococci (CNS). While the PCR product of femA (509 bp) or femB (651 bp) was obtained from almost all the S. aureus strains except for five oxacillin-resistant strains (2'5%), neither of these genes were detected in CNS. Therefore, the detection offemA and femB together with mecA by PCR was considered to be a more reliable indicator to identify MRSA by differentiating it from mecA-positive CNS than single detection of mecA.

Three rare human G12 strains were detected from diarrheic clinical samples of children (<8 months of age) in Calcutta during a routine surveillance study of rotaviral diarrhea in India. The VP7 genes of G12 strains and their products showed maximum homology (97 to 99% at the nucleotide level and 98% at the amino acid level, respectively) with those of two recently reported G12 strains (from the United States and Thailand) but lesser homology with those of prototype G12 strain L26.Rotaviruses are the major cause of acute gastroenteritis in infants and young of a wide variety of mammalian and avian species (11). In developing countries, approximately 130 million infants are infected with rotaviruses and there are 800,000 annual deaths (5, 12). In group A rotaviruses, 15 VP7 G serotypes and 21 VP4 P genotypes have been reported for humans, animals, and birds (17). Though 10 G types and 10 P types from humans have been reported (15), clinically and epidemiologically the most important strains belong to the G1 to G4 serotypes with P[8] and P[4] genotypes (6,14). On the other hand, some unusual types (G5, G8, and G9) and rare combinations of G and P types from different countries have also been reported (1,2,3,4,13,16,20). The human rotavirus G12 strains (L26 and L27) were first detected from Philippines in 1990 (19). After more than a decade, in 2002, human G12 strains Se585 (9) and T152 (15) from the United States and Thailand, respectively, were reported. In this study, we report the detection of three rare G12 strains of human rotaviruses in India.As part of a routine surveillance study for diarrheal diseases, stool samples were collected from children below 4 years of age from B. C. Roy Children's Hospital and also from patients of all age groups admitted to the Infectious Diseases Hospital, Calcutta, India, in 2001. Stool samples were also collected from children with diarrhea from Assam Medical College, Dibrugarh, Assam, India; Capital Hospital and Municipality Hospital, Bhubaneshwar, Orissa, eastern India; and Post-Graduate Institute of Medical Education and Research, Chandigarh, northern India. A total of 454 samples were screened for rotaviruses by RNA electrophoresis as described earlier by Herring et al. (10). The fecal specimens were processed for extraction of rotavirus double-stranded RNA suitable for reverse transcription and amplification of the VP4 and VP7 genes as described previously (4). G and P typing of positive samples was carried out by nested and multiplex PCR with consensus and type-specific primers as described previously (4,7,8,18,21). The amplified products were purified with either the QIAquick gel extraction kit or the PCR purification kit (Qiagen, GmBH) in accordance with the manufacturer's instructions. Direct sequencing was carried out by using ABI PRISM BigDye terminator cycle sequencing kits (Applied Biosystems) with an automated DNA sequencer, the ABI PRISM 310 genetic analyzer (Applied Biosystems). The sequencing of the VP7 genes of all three isolates was repeated three times to...

We developed a loop-mediated isothermal amplification (LAMP) method to detect Bordetella pertussis infection. This LAMP assay detected B. pertussis with high sensitivity, but not other Bordetella species. Among nasopharyngeal swab samples from subjects with suspected pertussis, LAMP results showed a high level of agreement with results of conventional PCR. This method is a rapid, sensitive, and specific method for diagnosis of B. pertussis infection even in clinical laboratories with no specific equipment.

Long electropherotype with Subgroup I specificity is a common feature of animal rotaviruses. In an epidemic of infantile gastroenteritis in Manipur, India, long but SG I strains predominated in the outbreak in the year 1987-88. One such strain isolated from that region, following the outbreak had G9P [19] specificity. As this is a rare combination, the gene sequences encoding VP4, VP6, VP7, NSP1, NSP2, NSP3, NSP4 and NSP5 of this strain were analyzed. All these genes except VP7 were closely related to porcine rotaviruses (95-99% identity at amino acid level) and clustered with the porcine strains in phylogenetic analysis. In addition, it had subgroup I nature and belonged to NSP4 genotype B which is characteristic of animal rotaviruses. This is the first report of a rotavirus with VP6 and NSP4, two crucial proteins thought to be involved in host range restriction and pathogenicity, were of porcine origin and caused diarrhoea in a human host. Among the genes of this strain sequenced so far, only VP7 had highest identity to human strains at amino acid level. This study suggests reassortment may be occurring between human and other animal strains and some of the reassortant viruses may be virulent to humans.

We have sequenced gene 5 encoding NSP1 for three human, two porcine, two bovine, one feline, and five equine rotavirus strains, and compared the nucleotide and deduced amino acid sequences with the published sequences for other various strains. Subgroup I human strains L26, 69M, and DS-1 were found to have a similar NSP1 sequence despite their different G serotypes, VP4 genotypes, and RNA patterns. The NSP1 sequence of the human strain K8 showed a high degree of homology to those of porcine strains OSU and YM. A high degree of homology was found among three equine strains (H2, FI-14, and FI23), but they differed from the other equine strains L338 and H1. The strain H1 was similar to the porcine strains. The feline strain Cat2 showed a high homology to bovine strains UK, RF, and A44. Thus, species-specific and interspecies relatedness of NSP1 sequences among human, porcine, bovine, feline and equine rotaviruses was found. Overall genomic relatedness of strains L26 and YM to various human and animal strains was also examined by RNA-RNA hybridization assay. The present and previous hybridization results showed that there is a good correlation in most strains between overall genomic property (or genogroup) and NSP1 sequence homology.