Do Plant Isolates Have a Role in Method Suitability and Growth Promotion Testing in the Microbiology Laboratory? Is It a Matter of Science versus Compliance?

In response to regulatory citations for not including plant isolates in method suitability and growth promotion testing of microbiological culture media, the authors make the case that the compendial designated cultures meet the requirements of the official tests and are sufficiently representative of the most frequently identified environmental isolates. It was our conclusion that this compliance request lacks scientific justification. The scope of this review was largely directed to the growth promotion and suitability testing requirements for USP <60>, <61>, <62>, and <71>. Other microbiological tests such as USP <51> Antimicrobial Effective Testing, media fill validation, and water and environmental monitoring are discussed.

Burkholderia cepacia: This Decision Is Overdue.

This is the third in a series of seven articles discussing the Recall Root Cause Research project conducted by the Division of Manufacturing and Product Quality, Center for Drug Evaluation and Research. This paper reviews the regulatory and scientific impact of a common and recurring opportunistic pathogen, Burkholderia cepacia. B. cepacia is comprised of closely related species called Burkholderia cepacia complex, which has contaminated many finished pharmaceutical products and environments used to manufacture pharmaceuticals. This review includes a brief perspective as described in several U.S. Food and Drug Administration (FDA) documents, and assesses root cause using product recall reports and FDA Establishment Inspection Reports. We identify several possible points of origin for microbial contamination. This discussion also includes concern with anomalies in test methods that may influence B. cepacia measurement. The issue of objectionable microorganisms and whether B. cepacia can readily be included in a compendial chapter is briefly discussed. Finally, this paper underscores that drugs contaminated with B. cepacia pose a serious threat to susceptible patients, particularly those with cystic fibrosis or who are otherwise immunocompromised. It is therefore important to prevent B. cepacia from contaminating pharmaceutical manufacturing environments, raw materials, and finished products. LAY ABSTRACT: Burkholderia cepacia is a species of bacterium that is commonly found in natural environments such as soil, water, rhizosphere and agriculture products. The species name represents a group of closely related organisms. These bacteria have contaminated many drug products and can create public health concerns. Pharmaceutical products that are contaminated with B. cepacia may pose serious consequences to vulnerable patients (e.g., compromised immune system). Preventing B. cepacia contamination in drugs by addressing the potential sources of this bacteria in a drug manufacturing operation is an important public health goal. This review highlights potential sources of B. cepacia species as they relate to U.S. Food and Drug Administration findings recorded in data from Establishment Inspection Reports and Warning Letters.

Detection of extrahepatic hepatitis C virus replication by a novel, highly sensitive, single-tube nested polymerase chain reaction.

We established a cell culture system for the replication of hepatitis C virus (HCV) by using human T and B leukemia cell lines. These 2 cell lines were infected in vitro by using HCV-positive pooled patient serum samples. HCV RNA was extracted from infected cell lines at different times after infection, and a sequence of the virus 5' untranslated region was analyzed. Hepatitis C minus-strand RNA was detected in the infected cell lines by highly strand-specific rTth (recombinant Thermus thermophilus DNA polymerase)-based reverse transcription followed by a novel, highly sensitive, single-tube nested polymerase chain reaction (PCR) method. PCR products were analyzed by direct DNA sequencing. These results indicate that the HCV can replicate in T and B lymphocytes. This model should represent a valuable tool for the detailed study of the initial steps of the HCV replication cycle and for the evaluation of antiviral molecules.