Collaborative study for the calibration of a replacement International Standard for diphtheria toxoid for use in flocculation test.

We present the results of a collaborative study for the establishment of a replacement International Standard (IS) for diphtheria toxoid for use in flocculation test and its calibration in Lf units. Calibration was performed using Ramon flocculation method, standardized using the 2nd IS. The candidate standard was assigned a unitage of 1870 Lf/ampoule based on results from 25 laboratories in 15 different countries and was established as the 3rd IS for diphtheria toxoid for use in flocculation test by the WHO Expert Committee on Biological Standardization (ECBS) in October 2015. The study also assessed the use of alternative methods for measuring Lf. Participants were asked to determine the Lf value of the candidate standard using an Enzyme Linked Immunosorbent Assay (ELISA) established at NIBSC, or other suitable in-house method. 10 laboratories performed ELISA according to the NIBSC protocol, 1 laboratory performed flocculation using laser-light scattering according to an in-house protocol, and 1 laboratory performed another in-house ELISA. Results suggest these methods may provide suitable alternatives to the Ramon flocculation test, subject to validation, and that the new standard could act as a suitable reference preparation in these methods.

Calibration of replacement international standards of diphtheria and tetanus toxoids for use in flocculation test.

The 1st International Reference Reagents (IRR) of Diphtheria and Tetanus Toxoids for Flocculation Test (DIFT and TEFT) were established by the WHO in 1988. These reagents are essential for the standardization of assays used to calculate Lf units of toxoids. Candidate replacement materials were provided by several European vaccine manufacturers and were formulated and freeze-dried at NIBSC. This paper provides a summary of the results of an international collaborative study including 18 laboratories from 16 countries, which examined the candidate replacement materials in a variety of methods. Materials 02/176 and 04/150 were proposed and adopted by the Expert Committee on Biological Standardization of WHO in October 2007 as 2nd WHO International Standards of Diphtheria and Tetanus Toxoid for use in Flocculation Test. The replacement standards were assigned the value of 1100 and 690Lf/ampoule, respectively, based on results of flocculation tests carried out using provided reagents. Material coded 02/176 fully complied with the WHO specifications for stability, residual moisture content, precision of fill and sterility. Stability of material coded 04/150 was slightly lower than expected but predictions were based only on 2-year data and were to be further monitored, post-adoption.

Comparison of maternal sera, cord blood, and neonatal sera for detecting presumptive congenital syphilis: relationship with maternal treatment.

The incidence of congenital syphilis has increased rapidly over the past few years. Most infected mothers and their newborns are asymptomatic at birth and diagnosis depends on serologic testing during pregnancy and at delivery. This study was initiated to compare maternal sera, cord blood, and neonatal sera for detecting presumptive congenital syphilis and to assess the role of maternal treatment (administration of penicillin to the mother at least 1 month before delivery) on the serologic results at the time of delivery. The serologic results from all live deliveries complicated by a positive maternal and/or neonatal test for syphilis during a 12-month period were compared using chi 2 analysis and multiple comparisons for proportions. Of 3306 livebirths, 73 (2.2%) were complicated by a positive maternal or neonatal serology. At delivery, the serologic test was positive in 68 (94%) of 72 maternal sera, 30 (50%) of 60 cord sera, and 43 (63%) of 68 neonatal sera. In the absence of maternal treatment, 95% of the maternal sera, 66% of the cord blood samples, and 86% of the neonatal sera were positive. If the mother had been treated, 94% of maternal sera, 36% of cord sera, and 39% of neonatal sera were positive. Cord blood and neonatal sera appear to be inferior to maternal sera for detecting prenatal exposure to syphilis. Cord serology is also inferior to neonatal serology at 2 to 3 days of age. The most effective way to identify newborns at risk for congenital syphilis is to obtain a maternal serologic diagnosis during pregnancy and to test maternal and neonatal sera at delivery.

The potential role of custody facilities in controlling sexually transmitted diseases.

BACKGROUND: The National Commission on Correctional Health Care recommends that medical screening, including tests for syphilis and other sexually transmitted diseases, be performed by the 14th day after initial booking procedures. Because the average length of stay in detention is usually less than 14 days, most detainees are not screened or treated for asymptomatic communicable diseases. METHODS: In order to determine the incidence and prevalence of syphilis among those booked through a large county jail, a rapid screening procedure to test newly incarcerated inmates and treat those infected was implemented over a 3-week period. RESULTS: Among 6309 detainees eligible for testing during the 3-week period of the study, 129 (2%) were diagnosed with syphilis. During the preceding month when only those reporting symptoms or high-risk behaviors were screened, 20 cases were diagnosed. The sensitivity and predictive value of self-reported risk factors for syphilis infection were low. Among the first 110 diagnosed cases, 17 (15.5%) were released within 3 days and 86 (78%) were released within 6 days. CONCLUSIONS: If screening and treatment for syphilis are delayed until the 14th day after booking, the majority of infected inmates will be released prior to evaluation and treatment. Rapid screening and treatment for selected sexually transmitted diseases are likely to be an effective public health measure when implemented in custody settings.

[Cytotoxicity of ferric chloride used as an adjuvant of organic acid precipitation]. / La cytotoxicité du chlorure ferrique utilisé comme adjuvant de la précipitation organique acide.

The objective of this study was the amelioration of the organic acid precipitation technique currently used as a secondary stage for viruses concentration using ferric chloride (FeCl3) as an adjuvant. To carry out this work successfully, we have studied the cytotoxicity effect of FeCl3 towards cell lines usually used for the investigation of enteric viruses. Two kinds of african green monkey kidney cells, MA104 and BGM, were used as a cellular model. The results showed that optimum concentration of FeCl3 for either BGM or MA104 cell survival and organic acid precipitation varies from 0.5 to 1 mM. The cytotoxicity test used in this work was simple, easy to realize and gives an appreciable information about the cytotoxicity dose of a given biological and chemical product.

[A method for recovery of enteroviruses from milk]. / Une méthode de recherche des entérovirus dans le lait.

A method of detection of enteric viruses in milk was studied. The high protein content of milk and the protein nature of enterovirus allowed the detection of these viruses using the organic acid flocculation method. The poliovirus type 1 (Mahoney strains) and the E.C.H.O.1 isolated from the environment were used as virus model and were inoculated to creamed, half-creamed and whole UHT commercialized milk. The method consists on a milk sample clarification with acid precipitation and centrifugation. The clarified extract is reduced to a final volume of 10 to 15 ml after addition of beef extract powder and protein precipitation. This technique allows the recovery of 26 to 36% of poliovirus type 1 and 10 to 46% of E.C.H.O.1 viruses. In this work, the ferric chloride (FeCl3), added in 0.5 to 1 mM final concentration, was used as an adjuvant for the organic acid precipitation.

Pretest temperature effects on CAP syphilis serology survey samples.

The College of American Pathologists syphilis serology survey participants periodically question survey-sample stability when the samples have possibly been exposed to temperature extremes during shipment. In this study it was demonstrated that short-term pretest incubations at temperatures above 58 degrees C (136.4 degrees F) usually cause a reduction in both rapid plasma reagin and FTA-ABS reactivity. No reactivity changes occurred in either test with short-term exposures to temperatures between -80 degrees C and 58 degrees C.