Standardization of fungal polymerase chain reaction for the early diagnosis of invasive fungal infection.

Background: An early initiation of antifungal therapy in invasive fungal infections (IFIs) is critical in reducing the high mortality rate. Current diagnosis of fungal infection relies on microscopy, culture, antigen, antibody specific tests and histological diagnosis. However, these tests either lack sensitivity or specificity. There is thus the need for a rapid, specific and accurate diagnostic method. Objective: The aim of our study was to establish PCR for the rapid detection of Candida and Aspergillus species in clinical specimens with improved sensitivity and specificity. Materials and Methods: A total of 71 proven cases of IFI (confirmed by culture) were collected. A total of 15 healthy, 15 patients suffering from bacterial sepsis and 15 patients with HIV, HBV viral infections were included as controls. Clinical specimens were subjected to a standardized nested amplification to produce Round I (504 bp) and Round II (150 bp) amplicons. Restriction digestion was performed on these products for further identification. Results: Analytical sensitivity was determined using 10 6 -10 CFU/ml of cell suspension. The lower detection limit of the assay was 10 CFU/ml of blood. This test was 100% sensitive and specific with a positive predictive value of 100% and a negative predictive value of 96.7%. Conclusion: The assay was found to be effective for the rapid detection of Candida and Aspergillus in clinical specimens.

Evaluation of BioFM liquid medium for culture of cerebrospinal fluid in tuberculous meningitis to identify Mycobacterium tuberculosis.

The present study was designed to evaluate the sensitivity and specificity of liquid culture medium (BioFM broth) for the diagnosis of tuberculous meningitis (TBM) in cerebrospinal fluid (CSF). CSF samples from 200 patients (TBM group = 150 and non-TBM group = 50) were tested for culture of Mycobacterium tuberculosis in BioFM liquid culture medium. Out of 150 TBM cases, 120 were found to be culture positive, indicating a sensitivity of 80% in BioFM broth within 2-3 weeks of inoculation. Positive cultures were also observed for CSF from 32 (64%) out of 50 non-TBM patients in BioFM liquid culture medium within 4 days of sample inoculation. Therefore, according to our study, BioFM broth system yielded 80% sensitivity [95% confidence interval (CI): 67-93%] and 36% specificity (95% CI: 57-98%) for TBM diagnosis. Our results indicate that although BioFM broth allows the detection of positive cultures within a shorter time, it has a high potential for contamination or for the coexistence of M. tuberculosis and non-tuberculous meningitis (NTM). This coexistence may go undetected or potentially lead to erroneous reporting of results.

Patent ductus arteriosus with a left to right shunt in a 64-year-old asymptomatic male.

We report here the unusual case of a 64 year old male, who was incidentally found to have a patent ductus arteriosus (PDA). The patient was asymptomatic. Echocardiography revealed a moderate sized PDA with normal left ventricular function and mild pulmonary hypertension. As the patient was asymptomatic and was not willing for any form of intervention, he was discharged and is currently under close follow-up. To the best of our knowledge this is the oldest reported case of a PDA in Indian literature.

Acute renal tubular dysfunction in association with Salmonella enteritidis.

A thirteen-year-old boy presented with acute renal tubular dysfunction after an infection with salmonella enteritidis. The child recovered following treatment with ciprofloxacin for a week.

Follow up of 318 cases of diabetes mellitus.

Ignorance, apathy, desire to get free advice, investigation and treatment is prevalent in diabetics. Most diabetics (69.63%) are uncontrolled whether on diet, single oral drug or combination of oral drugs or insulin. Ischemic heart disease was commonest complication. Neuritis was present in the one fourth of the followed up cases and was more prevalent in uncontrolled cases. Hypertension increases with the duration of diabetes and was twice more prevalent after duration of more than 5 years. Eye changes were present in about 50% of the people examined. Abnormality increases with the duration of diabetes.

Effect of azadirachtin on Mesocyclops leuckarti sensu lato, vector of dracunculiasis.

Extension or otherwise of characteristic insect development inhibitory effects of the neem sesquiterpenoid azadirachtin have been examined on eggs and nauplii of the crustacean cyclops, M. leuckarti sensu lato, a major vector of guineaworm disease at 0.1 ppm and above doses. Azadirachtin reduced survival of early nauplii (N1 to N3) at 0.1 ppm and above doses. No such effect was observed when eggs and late nauplii (N4 to N5) were exposed. Azadirachtin did not affect the moult cycle when late nauplii (N4 to N5) were exposed continuously till adulthood. At all concentrations tested, adult cyclops exhibited normal mating.

Neonatal osteomyelitis and septic arthritis.

The records of seventeen neonates (13 males and 4 females) with osteomyelitis and septic arthritis, who were admitted over a 3 year period, were reviewed. Neither prematurity nor low birth weight seemed to be a predisposing factor. The majority of the neonates (88.2%) had some antecedent illness or were subjected to certain procedures in the neonatal period. Paucity of movement of limb, fever and local swelling of the joint were the most frequent presenting symptoms. Deep soft tissue swelling was the frequently noted radiographic change (58.8%). Bone scintigraphy was abnormal in 7 out of 11 cases. Of the 21 sites involved, femur and humerus accounted for 76.1% of cases. Four of seventeen neonates had multiple bone involvement. Arthritis was present in fifteen neonates. Of the organisms isolated, Staphylococcus aureus was the commonest.