Distribution of Chlamydia trachomatis omp A genotypes in patients attending a sexually transmitted disease outpatient clinic in New Delhi, India

Background & objectives: Limited data are available on the typing of Chlamydia trachomatis in India. Serovars D to K of C. trachomatis are chiefly responsible for urogenital infections. Thus, this study was conducted to determine the distribution of C. trachomatis serovars in patients with urogenital infections and to characterize omp A gene of the detected C. trachomatis isolates by sequence analysis. Presence of other co-infections was also evaluated. Methods: Endocervical swabs were collected from 324 women and urethral swabs/urine were collected from 193 men attending the sexually transmitted diseases outpatient clinic. The samples were screened for C. trachomatis by cryptic plasmid PCR and omp A gene PCR. Genotyping was performed by PCR-restriction fragment length polymorphism (RFLP) and sequencing of the omp A gene. Samples were screened for genital mycoplasmas, Neisseria gonorrhoeae, Treponema pallidum and human immunodeficiency virus (HIV). Results: C. trachomatis was found in 15.0 per cent men and 10.8 per cent women. Serovar D was the most prevalent followed by serovars E, F, I and G. Twenty two C. trachomatis isolates were selected for omp A gene sequencing. No mixed infection was found. Variability in omp A sequences was seen in 31.8 per cent cases. Both PCR-RFLP and omp A gene sequencing showed concordant results. The presence of Ureaplasma spp. and Mycoplasma hominis was observed in 18.7 and 9.5 per cent patients, respectively. Co-infection of C. trachomatis was significantly associated with Ureaplasma urealyticum and HIV. Interpretation & conclusions: The high occurence of C. trachomatis infections warrants its screening in addition to other sexually transmitted infections namely U. urealyticum and HIV. Genotyping of the omp A gene may provide additional information for vaccine development.

Sonication of antibiotic loaded cement spacers: A valuable technique for detection of infection persistence in two-stage revision for infected joint arthroplasty

We evaluated the diagnostic utility of sonication of antibiotic loaded cement spacers comparing with periprosthetic tissue cultures for the detection of persisting infection in 14 patients undergoing staged procedures. Sonication improved microbial detection of intraoperative cultures from 14.2% to 28.5% (P = 0.481). Routine sonication of spacers is recommended.

Prosthetic joint infection: A major threat to successful total joint arthroplasty

Total joint arthroplasty (TJA) is one of the most common and reliable orthopaedic procedures that has significantly improved the quality of life of patients with degenerative joint diseases. Following the increase in the ageing population, availability of trained orthopaedic surgeons and advances in implantation procedures, demand for TJA both globally and in India is significantly increasing. Though TJA is one of the most cost-successful orthopaedic procedures, prosthetic joint infection (PJI) is one of the major complications of joint arthroplasty. Accurate diagnosis of PJI is challenging. Since total hip and knee arthroplasties comprises the majority of TJAs, this review focuses on the current understanding of incidence, risk factors, pathogenesis, causative microorganisms, diagnosis, treatment and prevention of PJI related to these two procedures.

Herpes simplex virus type 2 and cytomegalovirus perigenital ulcer in an HIV infected woman

We report a case of mucocutaneous Herpes Simplex Virus (HSV)-2 and Cytomegalovirus (CMV) infection in a 39-year-old female with acquired immunodeficiency syndrome, who presented with a perigenital ulcer. The patient was receiving antiretroviral treatment (ART) for 3 months before presentation. Scraping from the perigenital ulcer was positive for HSV-2 and Treponema pallidum using polymerase chain reactions (PCR). The extent and duration of the lesions led us to consider the possibility of coinfection with CMV. The patient also tested positive for CMV by PCR. On subsequent follow-up after 8 weeks, the genital lesions had healed completely. This is possibly ascribable to the ART, which led to significant immune reconstitution.

Sexually transmitted infections: Need for extragenital screening

Extragenital infections can occur concurrently with simultaneous urogenital infections. Extragenital sites are believed to serve as hidden reservoirs and play a critical role in their transmission. The etiological relationship of the most widespread Sexually transmitted diseases (STD) pathogen to reproductive tract has long been established, but the distribution to extragenital sites appears to be infrequent and its correlation with the sexual practice still requires to be investigated. Optimal-screening strategies for extragenital infections are largely unknown. However, there is a lack of data on clinical outcomes and optimal treatment regimens for rectal and pharyngeal extragenital infections. Further studies are needed in settings other than reproductive health and STD clinics, especially in primary care clinics and resource-limited settings.

Application of real‑time quantitative polymerase chain reaction assay to detect Legionella pneumophila in patients of community‑acquired pneumonia in a tertiary care hospital.

Legionella pneumophila is one of the important pathogen responsible for community –acquired pneumonia attributing for 1-5% of cases. Since early and accurate therapy reduces mortality, rapid and reliable diagnostic methods are needed. A total of 134 samples of blood, urine and respiratory tract fluids were collected. Blood was tested for IgG, IgM and IgA antibodies using commercially available kits. A total of 8 (6%) samples were found to be positive for L. pneumophila by quantitative reverse transcription polymerase chain reaction (qRT‑PCR), compared to conventional PCR where 6 (4.4%) samples were positive. Serology was positive in a total of 32 (23%) cases though only 3 (2.2%) of the PCR‑positive cases were positive by serology as well. These results suggest that real‑time PCR can detect Legionella infection early in the course of the disease before serological response develops.

Coagulase‑negative staphylococci causing blood stream infection at an Indian tertiary care hospital: Prevalence, antimicrobial resistance and molecular characterisation.

Introduction: Recent years have seen a rise of coagulase‑negative staphylococci (CoNS) from common contaminants to agents of nosocomial blood stream infections (BSI’s). Molecular typing and establishing a correlation with antibiotic resistance is essential particularly in countries like India where genotyping studies for drug‑resistant CoNS are sparse. Methods: A prospective study was done over 18 months, wherein 42,693 blood samples were received, and 59 patients with BSI due to CoNS were evaluated. The isolates recovered were identified by a biochemical test panel and matrix‑assisted laser desorption ionization – time of flight mass spectrometry followed by antimicrobial susceptibility testing by Kirby–Baur disc diffusion method and E‑test strips. Staphylococcal chromosomal cassette mec (SCCmec) element was characterised by multiplex polymerase chain reaction for all methicillin‑resistant (MR) isolates. Results: The majority of CoNS isolated were constituted by Staphylococcus haemolyticus (47.5%) followed by Staphylococcus epidermidis (33.9%), Staphylococcus hominis (11.86%), Staphylococcus cohnii (5.08%) and Staphylococcus warneri (1.69%). Among all isolates 57.6% were MR with statistically significant higher resistance versus methicillin sensitive‑CoNS. This difference was significant for erythromycin (76% vs. 44%, P = 0.011), rifampicin (50% vs. 12%, P = 0.002) and amikacin (26.5% vs. 4%, P = 0.023), ciprofloxacin (64.7% vs. 20%, P = 0.001) and cotrimoxazole (55.9% vs. 20%, P = 0.006). SCCmec type I was predominant (61.8%, P = 0.028) and exhibited multidrug resistance (76.2%). Coexistence of SCCmec type I and III was seen in 8.82% MR isolates. Conclusion: CoNS exhibit high antimicrobial resistance thereby limiting treatment options. The presence of new variants of SCCmec type in hospital‑acquired CoNS may predict the antibiotic resistance pattern. This is the first evaluation of the molecular epidemiology of CoNS causing BSI from India and can serve as a guide in the formulation of hospital infection control and treatment guidelines.

The changing face of community-acquired methicillin-resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is an important cause of infection, both in hospitalised patients with significant healthcare exposure and in patients without healthcare risk factors. Community-acquired methicillinresistant S. aureus (CA-MRSA) are known for their rapid community transmission and propensity to cause aggressive skin and soft tissue infections and community-acquired pneumonia. The distinction between the healthcare-associated (HA)-MRSA and CA-MRSA is gradually fading owing to the acquisition of multiple virulence factors and genetic elements. The movement of CA-MRSA strains into the nosocomial setting limits the utility of using clinical risk factors alone to designate community or HA status. Identification of unique genetic characteristics and genotyping are valuable tools for MRSA epidemiological studies. Although the optimum pharmacotherapy for CA-MRSA infections has not been determined, many CA-MRSA strains remain broadly susceptible to several non-β-lactam antibacterial agents. This review aimed at illuminating the characteristic features of CA-MRSA, virulence factors, changing clinical settings and molecular epidemiology, insurgence into the hospital settings and therapy with drug resistance.

Prosthetic joint infection due to Lysobacter thermophilus diagnosed by 16S rRNA gene sequencing.

We report the first case of prosthetic joint infection caused by Lysobacter thermophilus which was identified by 16S rRNA gene sequencing. Removal of prosthesis followed by antibiotic treatment resulted in good clinical outcome. This case illustrates the use of molecular diagnostics to detect uncommon organisms in suspected prosthetic infections.

Ureaplasma: Current perspectives.

Ureaplasma species are the most prevalent genital Mycoplasma isolated from the urogenital tract of both men and women. Ureaplasma has 14 known serotypes and is divided into two biovars‑ Ureaplasma parvum and Ureaplasma urealyticum. The organism has several genes coding for surface proteins, the most important being the gene encoding the Multiple Banded Antigen (MBA). The C‑terminal domain of MBA is antigenic and elicits a host antibody response. Other virulence factors include phospholipases A and C, IgA protease and urease. Besides genital tract infections and infertility, Ureaplasma is also associated with adverse pregnancy outcomes and diseases in the newborn (chronic lung disease and retinopathy of prematurity). Infection produces cytokines in the amniotic fluid which initiates preterm labour. They have also been reported from renal stone and suppurative arthritis. Genital infections have also been reported with an increasing frequency in HIV‑infected patients. Ureaplasma may be a candidate ‘co factor’ in the pathogenesis of AIDS. Culture and polymerase chain reaction (PCR) are the mainstay of diagnosis. Commercial assays are available with improved turnaround time. Micro broth dilution is routinely used to test antimicrobial susceptibility of isolates. The organisms are tested against azithromycin, josamycin, ofloxacin and doxycycline. Resistance to macrolides, tetracyclines and fluoroquinolones have been reported. The susceptibility pattern also varies among the biovars with biovar 2 maintaining higher sensitivity rates. Prompt diagnosis and initiation of appropriate antibiotic therapy is essential to prevent long term complications of Ureaplasma infections. After surveying PubMed literature using the terms ‘Ureaplasma’, ‘Ureaplasma urealyticum’ and ‘Ureaplasma parvum’, relevant literature were selected to provide a concise review on the recent developments.

Experimental and Clinical Evaluation of Nootropic Activity of Bacopa monniera Linn. (Brahmi).

Bacopa monniera Linn. (Brahmi) is an annual creeper belonging to family Scrophulariaceae and growing all over the Indian sub-continent in marshy areas. It is a major Medhya Rasayana used in Ayurveda for treatment of memory disorders. Large number of saponins and glycosides has been isolated from the plant. Most of the experimental and clinical studies have been done with crude extracts or standardized preparation of the two active saponins Bacosides A and B. Extracts or saponin mixture facilitate learning, improve consolidation of learned behavior and delay extinction in several models of learnt behavior in normal rats and mice as well as in chemically induced or transgenic models of Alzheimer's disease. They also prevent or reverse amnesia produced by drugs, stress or ischemic hypoxia. Other CNS effects include anti-anxiety, anti-convulsant and analgesic activity. Several mechanisms have been proposed to explain the mechanism of these CNS effects. Extracts as well as the bacoside preparation have been found safe and well tolerated in healthy volunteers in single dose or chronic administration for several weeks in a number of double blind placebo controlled studies in India and abroad. Chronic administration significantly improved information processing, learning and memory consolidation. It was found more effective than caffeine in a comparative study. Double blind placebo controlled studies with bacoside preparation have demonstrated beneficial effects and safety in elderly patients with Age Related Memory Impairment and in children with Attention Deficit Memory Disorder. It has also been found useful in anxiety neurosis, epilepsy and sleep disturbances in post menopausal women. The standardized preparation is marketed as a prescription drug after having obtained the necessary regulatory approval in India, Australia, New Zealand and South Africa and as an OTC product in several other south east Asian and African countries.

Bacteriological and antimicrobial susceptibility profile of soft tissue infections from Northern India.

BACKGROUND: Soft tissue infections require a judicious combination of antimicrobial therapy in addition to surgical debridement to limit tissue loss and preserve life. AIMS: To study the bacterial etiology of soft tissue infections and their antimicrobial susceptibility pattern. SETTINGS AND DESIGN: A single institutional retrospective study of one year duration from January to December 2002. MATERIALS AND METHODS: A total of 5039 consecutive pus samples received in the Bacteriology Laboratory was analyzed from the records. RESULTS: There were a total of 2783 bacterial isolates comprising of 1504 gram negative bacilli and 1279 gram positive cocci. Staphylococcus aureus was the commonest isolate followed by Escherichia coli and Pseudomonas species. Methicillin resistance in S. aureus was found to be 38.56%, high level aminoglycoside resistance was observed in 53.3% enterococci and 66.75% of the gram negative bacilli were extended spectrum beta-lactamase producers. Rifampicin and vancomycin showed best activity for S. aureus; for gram-negative bacilli, piperacillin-tazobactam combination showed best activity. CONCLUSIONS: Continuous monitoring of antimicrobial susceptibility pattern in individual settings together with their judicious use is emphasized to minimize emergence of drug resistant bacteria.

Antimicrobial resistance profile of nosocomial uropathogens in a tertiary care hospital.

A retrospective analysis was done of the resistance pattern of urinary tract pathogens isolated over a 4 months period in a tertiary care hospital. There were a total of 871 clinical isolates comprising of 793 gram negative bacilli and 78 gram positive cocci obtained from 5477 consecutive urine samples. Extended spectrum beta lactamase production was observed in 71.5% of the gram negative bacilli; of these 6.18% were also inhibitor resistant. High level aminoglycoside resistance was observed in 70.17% of Enterococcus isolates while methicillin resistance was documented in 23.8% of the Staphylococcus isolates. A high level of resistance was also noted for ciprofloxacin. Multidrug resistance is a common problem in hospitals which emphasizes the need for judicious use of antimicrobial agents and their continuous in vitro monitoring.