A Blueprint for the Stabilization of Sub-Valent Alkaline Earth Complexes.

The study of sub-valent Group 2 chemistry is a relatively new research field, being established in 2007 with the report of the first Mg(I) dimers. These species are stabilized by the formation of a Mg-Mg covalent bond; however, the extension of this chemistry to heavier alkaline earth (AE) metals has been frustrated by significant synthetic challenges, primarily associated with the instability of heavy AE-AE interactions. Here we present a new blueprint for the stabilization of heavy AE(I) complexes, based upon the reduction of AE(II) precursors with planar coordination geometries. We report the synthesis and structural characterisation of homoleptic trigonal planar AE(II) complexes of the monodentate amides {N(SiMe3 )2 }- and {N(Mes)(SiMe3 )}- . DFT calculations showed that the LUMOs of these complexes all show some d-character for AE = Ca-Ba. DFT analysis of the square planar Sr(II) complex [Sr{N(SiMe3 )2 }(dioxane)2 ]∞ revealed analogous frontier orbital d-character. AE(I) complexes that could be accessed by reduction of these AE(II) precursors were modelled computationally, revealing exergonic formation in all cases. Crucially, NBO calculations show that some d-character is preserved in the SOMO of theoretical AE(I) products upon reduction, showing that d-orbitals could play a crucial role in achieving stable heavy AE(I) complexes.

Structural Investigation of Magnesium Complexes Supported by a Thiopyridyl Scorpionate Ligand.

Herein, we report the synthesis of a series of heteroleptic magnesium complexes stabilized with the scorpionate ligand tris(2-pyridylthio)methanide (Tptm). The compounds of the general formula [Mg(Tptm)(X)] (1-X; X = Cl, Br, I) were obtained via protonolysis reaction between the proligand and selected Grignard reagents. Attempts to isolate the potassium derivative K(Tptm) lead to decomposition of Tptm and formation of the alkene (C5H4N-S)2C=C(C5H4N-S)2, and this degradation was also modelled using DFT methods. Compound 1-I was treated with K(CH2Ph), affording the degradation product [Mg(Bptm)2] (2; Bptm = {CH(S-C5NH3)2}-). We analyzed and quantified the steric properties of the Tptm ligand using the structural information of the compounds obtained in this study paired with buried volume calculations, also adding the structural data of HTptm and its CF3-substituted congener (HTptmCF3). These studies highlight the highly flexible nature of this ligand scaffold and its ability to stabilize various coordination motifs and geometries, which is a highly desirable feature in the design of novel organometallic reagents and catalysts.

Synthesis, characterisation and reactivity of group 2 complexes with a thiopyridyl scorpionate ligand.

Herein we report the reactivity of the proligand tris(2-pyridylthio)methane (HTptm) with various Alkaline Earth (AE) reagents: (1) dialkylmagnesium reagents and (2) AE bis-amides (AE = Mg-Ba). Heteroleptic complexes of general formulae [Mg(Tptm)(R)] (R = Me, nBu; Tptm = {C(S-C5H4N)3}-) and [AE(Tptm)(N'')] (AE = Mg-Ba; N'' = {N(SiMe3)2}-) were targeted from the reaction of HTptm with R2Mg or [AE(N'')2]2. Reaction of the proligand with dialkylmagnesium reagents led to formation of [{Mg(κ3C,N,N-C{Bu}{S-C5H4N}2)(µ-S-C5H4N)}2] (1) and [{Mg(κ3C,N,N-C{Me}{S-C5H4N}2)(µ-OSiMe3)}2] (2) respectively, as a result of a novel transfer of an alkyl group onto the methanide carbon with concomitant C-S bond cleavage. However, reactivity of bis-amide precursors for Mg and Ca did afford the target species [AE(Tptm)(N'')] (3-AE; AE = Mg-Ca), although these proved susceptible to ligand degradation processes. DFT calculations show that alkyl transfer in the putative [Mg(Tptm)(nBu)] (1m') system and amide transfer in 3-Ca is a facile process that induces C-S bond cleavage in the Tptm ligand. 3-Mg and 3-Ca were also tested as catalysts for the hydrophosphination of selected alkenes and alkynes, including the first example of mono-hydrophosphination of 4-ethynylpyridine which was achieved with high conversions and excellent regio- and stereochemical control.

Insights into the Composition and Structural Chemistry of Gallium(I) Triflate.

"GaOTf" is a simple, convenient source of low-valent gallium for synthetic chemistry and catalysis. However, little is currently known about its composition or reactivity. In this work, 71 Ga NMR spectroscopy shows the presence of [Ga(arene)n ]+ salts on oxidation of Ga metal with AgOTf in arene solvents. However, a more complex picture of speciation is uncovered by X-ray diffraction studies. In all cases, mixed-valence compounds containing Ga-arene and Ga-OTf coordination motifs, in addition to an unusual "naked" [Ga]+ ion, are found. Addition of 18-crown-6 allows for the isolation of a discrete GaI crown complex. Evidence of a potential intermediate in the formation of "GaOTf" has been isolated in the form of the bimetallic silver(I)/gallium(I) cluster anion [Ag4 {Ga(OTf)3 }4 (µ-Ga)6 (OTf)4 ]2- .

HPV genotype prevalence in Australian women undergoing routine cervical screening by cytology status prior to implementation of an HPV vaccination program.

BACKGROUND: Data on the prevalence of cervical HPV genotypes in Australia by age and by grade of cytological abnormality are sparse. OBJECTIVE: Measure prevalence of HPV genotypes among 2620 Australian women by age and cytology status. STUDY DESIGN: Women presenting for routine Pap smear screening were recruited from diverse regions, including a significant sample of Indigenous women. DNA extracts prepared from Thinprep specimens were HPV genotyped by Roche LINEAR ARRAY HPV. RESULTS: HPV prevalence and genotype distribution were stratified by age (mean 32.6y) and Pap smear result (cytology normal in 86.7%). Overall HPV prevalence was 38.7% with high-risk HPV prevalence of 26.5%. Prevalence of HPV (66.3% in women<21y to 15.3% in women>40y), multiple HPV infection (45.5% in <21y to 5.8% in >40y) and vaccine-targeted genotypes (HPV 6/11/16/18) (34.1% in <21y to 2.4% in >40y) declined significantly with age. The six most common genotypes were: HPV 16 (8.3%), 51 (5.1%), 53 (4.7%), 62 (4.3%), 89 (3.9%) and 52 (3.8%). HR-HPV prevalence increased from 21.1% in women with normal cytology to 80.9% in those with cytologically predicted high-grade abnormalities (HGAs) (p<0.001). The most common genotypes in women with HGAs were HPV 16 (51.1%), 18 (14.9%), 52 (12.8%), 31 (10.6%), and 33 (10.6%): all HR-types. CONCLUSION: Pre-vaccination cross-sectional prevalence of HR-HPV infection was high in this sample of Australian women attending for screening. HPV 16 was the commonest high-risk type detected at all ages and cytological grades.

Rapid determination of lymphogranuloma venereum serovars of Chlamydia trachomatis by quantitative high-resolution melt analysis (HRMA).

A quantitative high-resolution melt analysis assay was developed to differentiate lymphogranuloma venereum-causing serovars of Chlamydia trachomatis (L1 to L3) from other C. trachomatis serovars (D to K). The detection limit of this assay is approximately 10 copies per reaction, comparable to the limits of other quantitative-PCR-based methods.

Comparison of PapType to Digene Hybrid Capture 2, Roche linear array, and Amplicor for detection of high-risk human papillomavirus genotypes in women with previous abnormal pap smears.

PapType human papillomavirus (HPV) assay was compared to Hybrid Capture 2 (HC2), Amplicor (Amp), and Linear Array (LA) HPV tests in 894 women undergoing management for a high-grade Pap smear abnormality. The sensitivity in detection of underlying high-grade histological diagnosis by PapType was 90.3% and by HC2 was 79.8%, while by Amp and LA it was 92.4% and 91.6%, respectively. The specificities were 52.5%, 55.3%, 49.4%, and 51.7% for PapType, HC2, Amp, and LA, respectively.

Human papillomavirus prevalence among indigenous and non-indigenous Australian women prior to a national HPV vaccination program.

BACKGROUND: Indigenous women in Australia have a disproportionate burden of cervical cancer despite a national cervical screening program. Prior to introduction of a national human papilloma virus (HPV) vaccination program, we determined HPV genotype prevalence by Indigenous status and residence in remote areas. METHODS: We recruited women aged 17 to 40 years presenting to community-based primary health services for routine Pap screening across Australia. A liquid-based cytology (LBC) cervical specimen was tested for HPV DNA using the AMPLICOR HPV-DNA test and a PGMY09/11-based HPV consensus PCR; positive specimens were typed by reverse hybridization. We calculated age-adjusted prevalence by weighting to relevant population data, and determined predictors of HPV-DNA positivity by age, Indigenous status and area of residence using logistic regression. RESULTS: Of 2152 women (655 Indigenous), prevalence of the high-risk HPV genotypes was similar for Indigenous and non-Indigenous women (HPV 16 was 9.4% and 10.5%, respectively; HPV 18 was 4.1% and 3.8%, respectively), and did not differ by age group. In younger age groups, the prevalence of other genotypes also did not differ, but in those aged 31 to 40 years, HPV prevalence was higher for Indigenous women (35% versus 22.5%; P < 0.001), specifically HPV clades α5 (OR = 2.1, 95% CI 1.1 to 4.3) and α7, excluding type 18 (OR 1.9, 95% CI 1.1 to 3.3). In multivariate analysis, detection of any HPV genotype was strongly associated with smoking and Pap-test abnormalities, with both risk factors more common among Indigenous women. CONCLUSION: Although we found no difference in the prevalence of HPV16/18 among Australian women by Indigenous status or, for Indigenous women, residence in remote regions, differences were found in the prevalence of risk factors and some other HPV genotypes. This reinforces the importance of cervical screening as a complement to vaccination for all women, and the value of baseline data on HPV genotype prevalence by Indigenous status and residence for the monitoring of vaccine impact.

Human papillomavirus genotype prevalence in cervical biopsies from women diagnosed with cervical intraepithelial neoplasia or cervical cancer in Fiji.

BACKGROUND: There is currently limited information about human papillomavirus (HPV) genotype distribution in women in the South Pacific region. This study's objective was to determine HPV genotypes present in cervical cancer (CC) and precancers (cervical intraepithelial lesion (CIN) 3) in Fiji. METHODS: Cross-sectional analysis evaluated archival CC and CIN3 biopsy samples from 296 women of Melanesian Fijian ethnicity (n=182, 61.5%) and Indo-Fijian ethnicity (n=114, 38.5%). HPV genotypes were evaluated using the INNO-LiPA assay in archival samples from CC (n=174) and CIN3 (n=122) among women in Fiji over a 5-year period from 2003 to 2007. RESULTS: Overall, 99% of the specimens tested were HPV DNA-positive for high-risk genotypes, with detection rates of 100%, 97.4% and 100% in CIN3, squamous cell carcinoma (SCC) and adenosquamous carcinoma biopsies, respectively. Genotypes 16 and 18 were the most common (77%), followed by HPV 31 (4.3%). Genotype HPV 16 was the most common identified (59%) in CIN3 specimens, followed by HPV 31 (9%) and HPV 52 (6.6%). Multiple genotypes were detected in 12.5-33.3% of specimens, depending on the pathology. CONCLUSION: These results indicated that the two most prevalent CC-associated HPV genotypes in Fiji parallel those described in other regions worldwide, with genotype variations thereafter. These data suggest that the currently available bivalent and quadrivalent HPV vaccines could potentially reduce cervical cancers in Fiji by over 80% and reduce precancers by at least 60%.

Clearance of human papillomavirus in women treated for cervical dysplasia.

OBJECTIVE: To estimate human papillomavirus (HPV) clearance of women with cervical abnormalities after treatment. METHODS: Women attending dysplasia clinics between 2001 and 2007 with a new diagnosis of high-grade dysplasia or persistent low-grade dysplasia requiring treatment by excision or laser ablation were invited to participate. Cervical cytology, histology of biopsies collected at colposcopy, and HPV DNA detection and genotyping of 37 HPV genotypes on specimens collected at treatment and subsequent routine visits were examined. A log-rank test was used to compare the survival distribution between groups. RESULTS: Of the 1,649 women eligible at treatment (baseline), 1,207 (73%) were included in the analysis; 96% (n=1,159) had three or more posttreatment visits. At baseline and the subsequent three follow-up visits, the prevalence of women with HPV DNA detected was 84%, 53% (on average, 6.3 months after baseline), 44% (on average, 15.7 months after baseline), and 45% (on average, 24.3 months after baseline). The median time to HPV clearance was approximately 6 months for either HPV 16 (n=387) or HPV 18 (n=96), irrespective of concurrent detection of other types. On average, HPV 16 or HPV 18 types cleared faster than other types (P<.001). This association remained significant after adjustment for age, preoperative histology, number of preoperative histology results, and treatment type. CONCLUSION: Clearance times of HPV 16 and HPV 18 infections were similar to each another but shorter than other HPV types. LEVEL OF EVIDENCE: III.

Chlamydia trachomatis genotypes among men who have sex with men in Australia.

BACKGROUND: Chlamydia trachomatis is a common bacterial sexually transmitted infection in men who have sex with men (MSM), although little is known about its distribution in Australian MSM communities. METHODS: From 2004 to 2008, 612 consecutive C. trachomatis positive anal swab and urine samples were collected for genotyping and quantification from MSM attending 2 sexual health centers (Melbourne and Sydney). RESULTS: The most common serovars detected were D (35.2%), G (32.7%), and J (17.7%), although these distributions changed significantly by year and city. C. trachomatis infections (2.8%) involved more than 1 serovar and only 1 lymphogranuloma venereum isolate was detected. The majority of serovar strains showed an identical omp1 genotype, with only 7.5% showing genotypic variability. Serovar G infections were not associated with overseas sexual activity; whilst individuals with serovar J were less likely to have had a prior C. trachomatis infection, and with serovar E were those who had prior C. trachomatis infection. Symptoms were present in 68% of urethral infections and 28% anal infections, and were associated with gonorrheal coinfection (13.8%), prior C. trachomatis infection (20.6%) and increasing age. A higher C. trachomatis load was identified in anal samples versus urine (1.48 × 10(4) genome copies/anal swab; 3.72 × 10(3) copies/mL urine) and no association was made to concentration including the presence of symptoms and prior C. trachomatis infection. CONCLUSIONS: This is the largest study of C. trachomatis serovars in MSM: it is the first to report C. trachomatis rectal loads, and provides an overview on C. trachomatis serovars and genotypic variants that circulate in Australian MSM communities.

Analysis of Newcastle disease virus quasispecies and factors affecting the emergence of virulent virus.

Genome sequence analysis of a number of avirulent field isolates of Newcastle disease virus revealed the presence of viruses (within their quasispecies) that contained virulent F0 sequences. Detection of these virulent sequences below the ~1% level, using standard cloning and sequence analysis, proved difficult, and thus a more sensitive reverse-transcription real-time PCR procedure was developed to detect both virulent and avirulent NDV F0 sequences. Reverse-transcription real-time PCR analysis of the quasispecies of a number of Newcastle disease virus field isolates, revealed variable ratios (approximately 1:4-1:4,000) of virulent to avirulent viral F0 sequences. Since the ratios of these sequences generally remained constant in the quasispecies population during replication, factors that could affect the balance of virulent to avirulent sequences during viral infection of birds were investigated. It was shown both in vitro and in vivo that virulent virus present in the quasispecies did not emerge from the "avirulent background" unless a direct selection pressure was placed on the quasispecies, either by growth conditions or by transient immunosuppression. The effect of a prior infection of the host by infectious bronchitis virus or infectious bursal disease virus on the subsequent emergence of virulent Newcastle disease virus was examined.

Development and evaluation of an ompA quantitative real-time PCR assay for Chlamydia trachomatis serovar determination.

Knowledge of circulating Chlamydia trachomatis serovars can be beneficial for sexual network surveillance, monitoring treatment success, and associating specific clinical manifestations. Typically, C. trachomatis serovars are predicted by nucleotide sequencing of four variable domains within the ompA gene. However, sequencing procedures can be labor-intensive, are not readily available, and can lack the capacity to identify multiple serovars. This study describes the development and evaluation of a quantitative real-time PCR (qPCR) test algorithm for the rapid prediction of C. trachomatis serovars, including ocular (A to C) and anogenital (D to L3) strains. This test comprises a primary qPCR to confirm C. trachomatis positivity and the phylogenetic group(s) present and a secondary set of qPCRs to determine specific serovars. Cell culture isolates from 15 prototypic C. trachomatis serovars were correctly identified using this assay, with no cross-reactivity observed among serovars or with other common pathogenic microorganisms. Five hundred clinical specimens (previously diagnosed as being C. trachomatis positive) were evaluated by qPCR, with their results compared to results obtained by conventional sequencing. The qPCR identified 88.9% (423/476) complete matches (95% confidence interval [CI], 86 to 92%) of serovars compared to the results obtained using the sequence-based approach. Among the anogenital specimens, 2.4% (12/494) (95% CI, 1.3 to 4.2%) contained multiple serovars, categorized as single-serovar infections by conventional sequencing. Overall, this test exhibited high discriminatory success for predicting C. trachomatis serovars, particularly among anogenital infections. This is the first report of a qPCR typing assay offering differentiation of C. trachomatis serovars associated with both anogenital and ocular diseases.

HPV genotype prevalence in women with abnormal pap smears in Melbourne, Australia.

Carcinoma of the cervix and its precursor, high-grade cervical intraepithelial neoplasia (CIN2/3), are associated with persistent oncogenic Human papillomavirus (HPV) infection, particularly HPV 16 and 18. HPV genotype distribution varies with severity of cervical disease, patient demographics such as age, as well as geographical location. In this study, HPV genotype prevalence was determined, using the Roche Linear Array genotyping test, among a cohort of 1,676 women being managed with ablative or excisional treatment following colposcopically directed biopsies, who were referred initially due to cytological abnormalities. HPV genotype prevalence, including presence of single and multiple infections was assessed against both histological diagnosis and age. Overall, 83.9% of women were identified as HPV positive, comprising of 32.2% single and 51.7% multiple HPV infections. Of those with an available histological diagnosis at time-of-treatment (n = 899), HPV positivity increased significantly with disease severity: 62.4% (normal), 77.6% (CIN1), 92.6% (CIN2), and 97.9% (> or =CIN3) (P < 0.006). Similarly, a significant increase in high-risk (HR) HPV detection was observed with severity of disease (P < 0.005). The five most prevalent genotypes were HPV 16 (35.1%), 31 (12.6%), 51 (11.1%), 52 (9.9%), and 18 (8.5%). HPV 16 was the only genotype to demonstrate a significant increase in prevalence with increasing severity of histological or cytological disease (P < 0.0001). Multiple HPV infections, including multiple HR-HPV infections, declined significantly with age (P < 0.02). These findings provide the largest dataset of HPV genotype prevalence rates within Australian women, though are not representative of the general population.

Effect of ThinPrep preparation on human papillomavirus detection and genotyping in rectal samples by PCR.

Specimen-to-specimen carryover during ThinPrep slide preparation was evaluated by comparing human papillomavirus genotypes detected prior and subsequent to the ThinPrep processing of 121 PreservCyt samples. Overall, 52 samples generated concordant genotypes and 38 had additional and 21 had fewer genotypes postprocessing. PreservCyt samples should be aliquoted for PCR testing prior to ThinPrep processing.

Validation of an automated detection platform for use with the roche linear array human papillomavirus genotyping test.

An automated platform (BeeBlot) was evaluated in parallel with the recommended protocol for the hybridization and detection steps of the Roche Linear Array human papillomavirus (HPV) genotyping test using DNA from 143 cervical specimens. Genotyping profiles showed 100% concordance between the methods, suggesting that automation could complement the Roche Linear Array for enhanced speed and reproducibility.

Absence of a Chlamydia trachomatis variant, harbouring a deletion in the cryptic plasmid, in clients of a sexually transmissible infection clinic and antenatal patients in Melbourne.

A Chlamydia trachomatis (CT) variant, harbouring a 377 bp deletion in the cryptic plasmid, recently identified in Europe, has caused false-negative reporting of CT infections by various assays. This report is aimed at identifying whether this variant is present among clients of a sexual health clinic, or antenatal screening patients in Melbourne. Two hundred CT-positive specimens (by BDProbeTec ET assay) from Melbourne Sexual Health Centre (August 2005-November 2006) were tested by COBAS TaqMan 48 PCR assay. Discrepancies were tested by an in-house real-time (Re-Ti) polymerase chain reaction (PCR) assay, amplifying a 274-bp region of the omp1 gene. Additionally, 1,071 consecutive specimens from antenatal screening patients at the Royal Women's Hospital (December 2006-April 2007) were tested by COBAS TaqMan 48 and omp1 Re-Ti PCR. The CT variant was not detected among the 200 CT-positive specimens (95% confidence interval 0-2.3%). Three tested CT-negative by COBAS TaqMan 48, omp1 Re-Ti PCR and CT mutant-specific PCR, suggesting sample degradation or differential assay sensitivity. Of the 1,071 antenatal screening specimens, 56 tested CT-positive and 1,015 CT-negative by COBAS TaqMan 48. All of the CT-negatives tested negative by omp1 Re-Ti PCR (95% confidence interval 0-0.5%), with 51 of 56 CT-positives testing positive. These findings show there were no CT variants among attendees of a Melbourne sexual health clinic, nor among antenatal screening patients. It is likely that the variant strain has not yet entered circulation in these populations. However, given the current upsurge in urogenital CT-infections, continued surveillance is necessary to ensure timely detection of this variant, should it be introduced into the population.

Development and validation of a real-time PCR assay specifically detecting human papillomavirus 52 using the Roche LightCycler 480 system.

The recently released Linear Array human papillomavirus (HPV) genotyping test (Roche Diagnostics) provides a standardized method for simultaneous detection of up to 37 individual HPV types. This test offers a rapid approach for detecting and longitudinal monitoring of patients infected with high-risk (HR) HPV. However, it cannot rule out HPV52 infections in the presence of carcinogenic HPV genotypes 33, 35 and/or 58. As such, often only a non-definitive result of HPV52 presence can be reported. This study describes the development of a real-time PCR assay using an HPV52-specific hydrolysis probe in conjunction with the newly released Roche LightCycler 480 system. HPV52 was readily detected among DNA extracts from samples previously identified with possible HPV52 by the linear array test. Specificity was analyzed using a panel of DNA extracts previously identified as containing single/multiple HPV types, with or without HPV52. This is a rapid and simple assay, which could be used as a supplementary test to the Linear Array, to verify the presence or absence of HPV52 among samples testing positive for either HPV33, 35, and/or 58. Laboratories already using the Linear Array HPV genotyping test could adopt this method once internally validated.

Comparison of the Digene Hybrid Capture 2 assay and Roche AMPLICOR and LINEAR ARRAY human papillomavirus (HPV) tests in detecting high-risk HPV genotypes in specimens from women with previous abnormal Pap smear results.

The development of cervical cancer is strongly associated with the presence of persistent high-risk (HR) human papillomavirus (HPV) infection. Recently, the commercially manufactured PCR-based Roche AMPLICOR (AMP) and LINEAR ARRAY (LA) HPV tests have become available for HPV detection. However, knowledge of their clinical performance compared to the U.S. Food and Drug Administration-approved Hybrid Capture 2 (HC2) assay is limited. This study evaluated the concordance between the HC2, AMP, and LA tests in detecting HR-HPV among a cohort of 1,679 women with previous abnormal Pap smear results. Overall, 1,393 specimens (81.3%) generated concordant results for HR-HPV presence or absence by the three assays. The concordance levels were substantial between the HC2 and AMP tests (84.4%, kappa = 0.6419) and between the HC2 and LA tests (84.0%, kappa = 0.6341) and nearly perfect between the AMP and LA tests (97.8%, kappa = 0.9441). HR-HPV prevalence, as detected by the AMP or LA tests, was significantly higher among women with cytological or histological high-grade disease (CIN2 or greater) than that detected by HC2 (P < 0.0001). The AMP and LA tests exhibited greater sensitivity, but lower specificity, than HC2 for detecting HR-HPV among this cohort of women with underlying cervical abnormalities, particularly among subjects with histologically proven high-grade disease. Both PCR-based HPV tests may be valuable in the management of care for women with underlying cervical abnormalities, in predicting treatment success, and in studying the clearance or acquisition of new infections.

Assessment of MagNA pure LC extraction system for detection of human papillomavirus (HPV) DNA in PreservCyt samples by the Roche AMPLICOR and LINEAR ARRAY HPV tests.

Roche Molecular Systems recently released two PCR-based assays, AMPLICOR and LINEAR ARRAY (LA), for the detection and genotyping, respectively, of human papillomaviruses (HPVs). The manual specimen processing method recommended for use with both assays, AmpliLute, can be time-consuming and labor-intensive and is open to potential specimen cross-contamination. We evaluated the Roche MagNA Pure LC (MP) as an alternative for specimen processing prior to use with either assay. DNA was extracted from cervical brushings, collected in PreservCyt media, by AmpliLute and MP using DNA-I and Total Nucleic Acid (TNA) kits, from 150 patients with histologically confirmed cervical abnormalities. DNA was amplified and detected by AMPLICOR and the LA HPV test. Concordances of 96.5% (139 of 144) (kappa=0.93) and 95.1% (135 of 142) (kappa=0.90) were generated by AMPLICOR when we compared DNA extracts from AmpliLute to MP DNA-I and TNA, respectively. The HPV genotype profiles were identical in 78.7 and 74.7% of samples between AmpliLute and DNA-I or TNA, respectively. To improve LA concordance, all 150 specimens were extracted by MP DNA-I protocol after the centrifugation of 1-ml PreservCyt samples. This modified approach improved HPV genotype concordance levels between AmpliLute and MP DNA-I to 88.0% (P=0.043) without affecting AMPLICOR sensitivity. Laboratories that have an automated MP extraction system would find this procedure more feasible and easier to handle than the recommended manual extraction method and could substitute such extractions for AMPLICOR and LA HPV tests once internally validated.