Evaluation of the indirect hemagglutination assay for diagnosis of acute leptospirosis in Hawaii.

Timely diagnosis of leptospirosis is important to ensure a favorable clinical outcome. The definitive serologic assay, the microscopic agglutination test (MAT), requires paired sera and is not useful for guiding early clinical management. The only screening test approved for use in the United States, the indirect hemagglutination assay (IHA), has not undergone extensive field evaluation. To assess the performance of the leptospirosis IHA in Hawaii, serum from patients evaluated for leptospirosis between 1992 and 1997 were tested with the IHA at the Hawaii State Laboratories Division and with the MAT at the Centers for Disease Control and Prevention. Leptospirosis was considered confirmed by a fourfold rise in MAT titer and/or a positive culture. A total of 92 (41%) of 226 specimens from 114 persons with confirmed leptospirosis were found positive by IHA. Only 18 (15%) of 119 specimens obtained within 14 days of onset were IHA positive, compared to 74 (69%) of 107 specimens collected more than 14 days after onset (P <0.001). Repeat testing ultimately resulted in 78 (68%) of the confirmed cases having at least one specimen found positive by IHA. Thirteen different presumptive infecting serogroups were identified among 251 specimens with an MAT titer of >/=200 and obtained from persons with confirmed or probable leptospirosis. Fifty (68%) of 73 specimens with Icterohaemorrhagiae as the presumptive infecting serogroup were found positive by IHA, compared to 44 (47%) of 93 specimens with Australis as the presumptive infecting serogroup (P, 0.01). The IHA test was positive for 3 (1%) of 236 specimens from 154 persons without leptospirosis. The sensitivity of the leptospirosis IHA in Hawaii was substantially below figures reported previously, particularly early in the course of illness, limiting its usefulness for diagnosing acute infection. Since the presumptive infecting serogroup affected IHA results and the prevalence of serovars varies with geography, the performance of the IHA should be assessed locally. More sensitive leptospirosis screening tests are needed in Hawaii.

Stimulatory effect of procaine on the growth of several microalgae and cyanobacteria.

Procaine has been used to stimulate plant growth and it has been noted that it also promotes growth of microorganisms. The effect of procaine hydrochloride concentration on the growth rates of several species of microalgae and cyanobacteria was studied under both photoautotropic and heterotrophic growth conditions. Procaine hydrochloride was added to cultures at concentrations over the range 0.01-1000 mg L(-1). A stimulating effect of procaine hydrochloride on photoautotrophic growth was observed for the cyanobacteria Anabaena cylindrica and Anabaena variabilis, and for the salt-tolerant green algae Dunaliella primolecta and Dunaliella parva. During active growth in batch culture an increase in growth rate (compared with control culture without procaine hydrochloride) of about 25% was observed at 0.1 mgL(-1) of procaine hydrochloride for A. cylindrica. However, procaine hydrochloride was toxic at concentrations of > 10 mgL(-1). Simultaneous administration of hydrolysis products of procaine, p-aminobenzoic acid and diethyl aminoethanol, in lieu of procaine hydrochloride, was as effective as procaine in stimulating growth of A. cylindrica. Heterotrophic growth of Chlorella ellipsoidea and Prototheca zopfii was not stimulated by procaine hydrochloride over the concentration range studied (0.1-10 mg L(-1)). The combined effects of procaine hydrochloride concentration and four other environmental factors (temperature, light intensity, CO2 concentration in the flushing gas and NaCl concentration) on growth rate of D. primolecta was modelled using both a neural network approach and a response surface method. These results indicate that procaine hydrochloride exerts different effects on the growth of microalgal and cyanobacterial cells as functions of dosage, species and culture conditions.

International multicenter evaluation of the clinical utility of a dipstick assay for detection of Leptospira-specific immunoglobulin M antibodies in human serum specimens.

We performed a multicenter evaluation of a robust and easily performed dipstick assay for the serodiagnosis of human leptospirosis. The assay is aimed at the detection of Leptospira-specific immunoglobulin M (IgM) antibodies. The study involved 2,665 serum samples collected from 2,057 patients with suspected leptospirosis in 12 countries on five continents with different levels of endemicity and different surveillance systems. The patients were grouped as laboratory-confirmed leptospirosis case patients and noncase patients based on the results of culturing and the microscopic agglutination test. Paired samples from 27.7% of the subjects were tested. Of the 485 case patients, 87.4% had a positive dipstick result for one or more samples. Of the 1,513 noncase patients, only 7.2% had a positive result. Whereas most (88.4%) of the positive samples from the case patients showed moderate to strong (2+ to 4+) staining in the dipstick assay, most (68.1%) of the positive samples from the noncase patients showed weak (1+) staining. The sensitivity of the dipstick assay increased from 60.1% for acute-phase serum samples to 87.4% for convalescent-phase samples. The specificities for these two groups of samples were 94.1 and 92.7%, respectively. The dipstick assay detected a broad variety of serogroups. The results of the dipstick assay were concordant (observed agreement, 93.2%; kappa value, 0.76) with the results of an enzyme-linked immunosorbent assay for the detection of specific IgM antibodies, a test which is often used in the laboratory diagnosis of current or recent leptospirosis. This study demonstrated that this easily performed dipstick assay is a valuable and useful test for the quick screening for leptospirosis; has a wide applicability in different countries with different degrees of endemicity; can be used at all levels of the health care system, including the field; and will be useful for detecting and monitoring outbreaks of leptospirosis.