Rapid diagnosis of disseminated Mycobacterium mucogenicum infection in formalin-fixed, paraffin-embedded specimen using nextgeneration sequencing: A case report

BACKGROUND Mycobacterium mucogenicum(M.mucogenicum)belongs to the group of rapidly growing Nontuberculous mycobacteria.This microorganism is associated with a wide spectrum of infectious diseases.Due to a low detection rate or the time required for conventional culture methodology,a rapid and broad-spectrum method is necessary to identify rare pathogens.CASE SUMMARY A 12-year-old immunocompetent girl presented with painful masses for five months.The first mass was found in the right upper quadrant of the abdomen,and was about 1 cm×1.5 cm in size,tough but pliable in texture,with an irregular margin and tenderness.An abscess gradually formed and ulcerated with suppuration of the mass.Three new masses appeared on the back one by one.Chest computed tomography showed patchy and streaky cloudy opacities in both lungs.Needle aspiration of the abscess was performed,but the smear and conventional culture were negative,and the pathological examination showed no pathogens.We then performed next-generation sequencing using a formalinfixed,paraffin-embedded specimen to identify the pathogen.A significantly high abundance of M.mucogenicum was detected.The patient’s abscesses gradually decreased in size,while inflammation in both lungs improved following 12-wk of treatment.No recurrence was observed four months after the end of the one-year treatment period.CONCLUSION Next-generation sequencing is a promising tool for the rapid and accurate diagnosis of rare pathogens,even when using a formalin-fixed,paraffin-embedded specimen.

RAPID DIAGNOSIS AND TREATMENT OF CHLAMYDIAL CONJUNCTIVITIS

During December 1989 to December 1992, conjunctival specimens from 63 patients with severe trachoma and 30 with acute follicular conjunctivitis at the eye clinic of Tong Ren Hospital in Beijing, were tested by using direct immunofluorescent technique and amplified enzyme-inked immunoassay (Micro Trak and IDEIA) for chlamydial antigenicity. Trachoma accounted for 97.6% while acute follicular conjunctivitis accounted for 2.4% of the positive cases. Micro Trak and IDEIA tests proved advantageous over the culture method for the detection of Chlamydia trachomatis. In our study, Ofloxacin eye ointment has been proved to be an alterative drug for severe trachoma with better curative effectiveness.

Rapid determination of lithium-ion battery degradation: High C-rate LAM and calculated limiting LLI

Herein,incremental capacity-differential voltage (IC-DV) at a high C-rate (HC) is used as a non-invasive diagnostic tool in lithium-ion batteries,which inevitably exhibit capacity fading caused by multiple mechanisms during charge/discharge cycling.Because battery degradation modes are complex,the simple output of capacity fading does not yield any useful data in that respect.Although IC and DV curves obtained under restricted conditions (<0.1C,25℃) were applied in non-invasive analysis for accurate observation of degradation symptoms,a facile,rapid diagnostic approach without intricate,complex calculations is critical in on-board applications.Herein,Li Ni_(0.5)Mn_(0.3)Co_(0.2)O_(2)(NMC532)/graphite pouch cells were cycled at 4 and 6C and the degradation characteristics,i.e.,loss of active materials (LAM) and loss of lithium inventory (LLI),were parameterized using the IC-DV curves.During the incremental current cycling,the initial steep LAM and LLI slopes underwent gradual transitions to gentle states and revealed the gap between low-and high-current measurements.A quantitative comparison of LAM at high and low C-rate showed that a IC;revealed the relative amount of available reaction region limited by cell polarization.However,this did not provide a direct relationship for estimating the LAM at a low C-rate.Conversely,the limiting LLI,which is calculated at a C-rate approaching 0,was obtained by extrapolating the LLI through more than two points measured at high C-rate,and therefore,the LLI at 0.1C was accurately determined using rapid cycling.

Accuracy of a new rapid diagnostic test for urinary antigen detection and assessment of drug treatment in opisthorchiasis

Background Screening for opisthorchiasis,a parasitic worm infection affecting many millions of people in Southeast Asia,has traditionally relied on faecal egg examination such as the formalin-ethyl acetate concentration technique(FECT)and Kato-Katz method.Although the urinary enzyme-linked immunosorbent assay(ELISA)has been used more recently,we developed a urinary antigen-based rapid diagnostic test(RDT)to simplify diagnosis and as a point-of-care testing(POCT)and field applications for surveillance and control of opisthorchiasis.Methods A urinaryOpisthorchis viverrini(OV)-RDT was developed using immunochromatographic methodology with a specific monoclonal antibody against OV.The diagnostic performance of the urinary OV-RDT was compared to that of quantitative faecal FECT and urinary antigen ELISA(n=493).Cross-reactivities of urinary OV-RDT with other helminthiases coexisted withO.viverrini were determined(n=96).A field trial in the application of urinary OV-RDT was compared with urinary antigen ELISA at baseline screening and assessment of drug treatment outcomes in opisthorchiasis(n=1629).The McNemar chi-square,Kruskal-Wallis and Cohen’s kappa coefficient(κ-value)tests were used for statistical analyses.Results Urinary OV-RDT had sensitivity of 94.2%and specificity of 93.2%,compared to faecal FECT.Urinary OV-RDT had high diagnostic agreement(Kappa=0.842-0.874,P<0.001)and quantitative correlation with urinary antigen ELISA(Kruskal-Wallis tests=316.2,P<0.0001)and faecal FECT(Kruskal-Wallis tests=362.3,P<0.0001).The positive rates by OV-RDT,ELISA and FECT were 48.9%,52.5%and 49.3%,respectively.Cross-reactions of urinary OV-RDT with other helminthiases were few(2%).Field trials of urinary OV-RDT yielded comparable prevalence ofO.viverrini between urinary OV-RDT(53.2%)and urinary antigen ELISA(54.0%).OV screening showed high diagnostic agreement(kappa>0.8,P<0.0001)between urinary OV-RDT and urinary antigen ELISA.The cure rates of opisthorchiasis at 1 month post-praziquantel treatment determined by urinary OV-RDT(86.6%)and urinary antigen ELISA(80.5%)were similar(P>0.05).Conclusions The urinary OV-RDT test has high potential as a new tool for screening and evaluating treatment outcomes in opisthorchiasis.The ease of sample collection and simplicity of urinary OV-RDT may facilitate mass screening,control and elimination of opisthorchiasis,thereby contributing to a reduction in the disease burden in Southeast Asia.

In-service Structural Health Monitoring of a Full-scale Composite Horizontal Tail

In-service structural health monitoring（SHM） technologies are critical for the utilization of composite aircraft structures. We developed a Lamb wave-based in-service SHM technology using built-in piezoelectric actuator/sensor networks to monitor delamination extension in a full-scale composite horizontal tail. The in-service SHM technology combine of damage rapid monitoring（DRM） stage and damage imaging diagnosis（DID） stage allows for real-time monitoring and long term tracking of the structural integrity of composite aircraft structures. DRM stage using spearman rank correlation coeffi cient was introduced to generate a damage index which can be used to monitor the trend of damage extension. The DID stage based on canonical correlation analysis aimed at intuitively highlighting structural damage regions in two-dimensional images. The DRM and DID stages were trialed by an in-service SHM experiment of CFRP T-joint. Finally, the detection capability of the in-service SHM technology was verified in the SHM experiment of a full-scale composite horizontal tail. Experimental results show that the rapid monitoring method effectively monitors the damage occurrence and extension tendency in real time; damage imaging diagnosis results are consistent with those from the failure model of the composite horizontal tail structure.

QF-PCR as a molecular-based method for autosomal aneuploidies detection

Objectives: The currently available methods for rapid prenatal diagnosis of common chromosomal aneuploidies are either Interphase-Fluorescence in Situ Hybridisation (I-FISH) or Quanti- tative Fluorescent Polymerase Chain Reaction (QF-PCR). QF-PCR represents a rapid, high throughput, cost-effective alternative for Interphase-FISH. The objective of the study was to evaluate the performance of QF-PCR, as a molecular-based technique for the detection of chromosome 21, 18 and 13 copy numbers. Study design: A retrospective cohort of 163 samples referred for screening of common chromosomal aneuploidies was blindly tested for chromosome 21, 18 and 13 copy numbers using QF-PCR and the results were compared with those of conventional cytogenetic analysis. Results: QF-PCR demonstrated optimal sensitivity and specificity (100%) for non mosaic trisomies. QF-PCR was able to consistently detect maternal cell contamination and mosaic trisomies when the trisomic cell line was present at an adequate level (23% or more). However, QF-PCR was unable to detect chromosomal rearrangements for which the primers were not designed. Conclusion: QF- PCR proved its superior performance as a molecular-based method for autosomal aneuploidy detection concerning both sensitivity and specificity.

Assessing the Diagnostic Accuracy of SD BIOLINE for Detection of Hepatitis C Virus:A Systematic Review and Meta-analysis

Hepatitis C virus(HCV)is a globally widespread ribonucleic acid virus that transmits through blood and sexual contact.Its morbidity and mortality are particularly higher in economically underdeveloped areas.Therefore,an economical and effective diagnostic method for detection of HCV is urgently needed.In this study,we evaluated the diagnostic accuracy of the SD BIOLINE rapid diagnostic test for HCV detection.We searched for studies related to SD BIOLINE and HCV in PubMed,Embase,Web of Science,and the Cochrane Library and then designed inclusion and exclusion criteria.After extracting valid data,the included literature was evaluated with the quality assessment tool Quality Assessment of Diagnostic Accuracy Studies.After our data analysis,the sensitivity,specificity,positive likelihood ratio,negative likelihood ratio,diagnostic accuracy,summary receiver operating characteristic curve,funnel plot,box plot,and Fagan plot of the diagnosticmethod were determined.Nine articles with nine sets of data were finally included.The sensitivity and specificity were 0.94 and 0.98,respectively,the positive likelihood ratio was 79.53,the negative likelihood ratio was 0.05,the diagnostic odds ratio was 1590.32,and the summary receiver operating characteristic curve was 0.9958.The SD BIOLINE test has the advantages of high sensitivity,high specificity,low cost,and easy operation for diagnosing HCV.Therefore,we recommend using SD BIOLINE for rapid and effective screening of HCV,which is especially applicable for economically underdeveloped areas.

Diagnostic Performance of the GenoType MTBDRplus and MTBDRs/Assays to Identify Tuberculosis Drug Resistance in Eastern China

Background： The WHO recently has recommended the GenoType MTBDRph/s version 1.0 and MTBDRs/version 1.0 assays for widespread use in countries endemic with drug-resistant tuberculosis. Despite this, these assays have rarely been evaluated in China, where the burden of drug-resistant tuberculosis is among the highest globally. Methods： Mycobacterium tuberculosis clinical isolates were obtained between January 2008 and December 2008. Isolates were tested for drug resistance against rifampicin （RFP） and isoniazid （INH） using the GenoType MTBDRplus assay and drug resistance against ethambutol （EMB）, ofloxacin （OFX）, and kanamycin （KM） using the Genotype MTBDILsl assay. These results were compared with conventional drug-susceptibility testing （DST）. Results： Readable results were obtained from 235 strains by GenoType MTBDRphts assay. Compared to DST, the sensitivity of GenoType MTBDRplus assay to detect RFP, INH, and multidrug resistance was 97.7%, 69.9%, and 69.8%, respectively, whereas the specificity for detecting RFP, INH, and multidrug resistance was 66.7%, 69.2%, and 76.8%, respectively. The sensitivity and specificity of the GenoType MTBDRsl assay were 90.9% and 95.2% for OFX, 77.8% and 99.5% for KM, 63.7% and 86.4% for EMB, respectively. Mutations in codon S531L of the rpoB gene and codon S315T1 ofKatG gene were dominated in multidrug-resistant tuberculosis （MDR-TB） strains. Conclusions： In combination with DST, application of the GenoType MTBDRplus and MTBDRsl assays may be a useful supplementary tool to allow a rapid and sale diagnosis of multidrug resistance and extensively drug-resistant tuberculosis.