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ABSTRACT Loop-mediated isothermal amplification (LAMP) is a simple and efficient nucleic acid amplification method for the rapid diagnosis of infectious diseases. This study assessed the performance of an in-house LAMP for tuberculosis (TB) diagnosis at a remote reference laboratory in the endemic setting of Thailand. As part of the routine service, 1,882 sputum samples were processed for mycobacterial culture in Lowenstein-Jensen and MGIT media. The DNA was extracted from the remaining decontaminated samples after the culture procedure for real-time polymerase chain reaction (PCR) analysis using Anyplex plus MTB/NTM detection kit. 785 (40.28%) were positive by mycobacterial culture. Of these, 222 DNA remnants were available and subjected to LAMP analysis. Based on culture as reference (Mycobacterium tuberculosis; MTB= 209/ non-tuberculous mycobacteria; NTM= 13), the overall sensitivity of LAMP and Anyplex plus assays for MTB detection were 89.95% (188/209; 95% confidential interval [CI]: 85.05-93.67%) and 96.65% (202/209; 95% CI: 93.22-98.64%), and the accuracy values were 88.74% (197/222; 95% CI: 83.83-92.58) and 96.40% (214/222; 93.02-98.43%), respectively. The sensitivity and accuracy of the in-house LAMP and the Anyplex plus real-time PCR assay were high in comparison to culture results. The high sensitivity and accuracy suggested that this in-house LAMP was promising and might be useful for early TB diagnosis.
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@#Abstract: Objective To analyze the laboratory microscopic re-examination results of malaria cases in Nantong of the National Notifiable Disease Report System from 2014 to 2021 by Nantong Malaria Diagnostic Reference Laboratory, so as to evaluate the malaria diagnosis ability of Nantong Malaria Diagnostic Reference Laboratory. Methods The blood smear and blood samples of malaria cases in Nantong from 2014 to 2021 of the National Notifiable Disease Report System were collected. Nantong Malaria Diagnostic Reference Laboratory and Jiangsu Institute of Parasitic Diseases carried out the re-examination of municipal and provincial laboratories, taking the results of provincial laboratory as the standard to compare and analyze the re-examination results of Nantong Malaria Diagnostic Reference Laboratory. Results From 2014 to 2021, the two-level laboratories in Nantong city and Jiangsu Province re-examined the blood samples of 297 malaria cases. The microscopic examination and PCR re-examination results at the provincial level were the same：292 positive cases and 5 negative cases. The qualitative coincidence rate between Nantong microscopic re-examination results and the provincial re-examination results was 100% (297/297), without misjudgment and omission. The coincidence rate of Plasmodium typing was 96.23% (281/292). The coincidence rate of P. falciparum, P. vivax, P. ovale and P. malaria were 99.57% (234/235), 62.50% (5/8), 89.47% (34/38) and 72.73% (8/11) respectively. The consistency test results showed that the Kappa value of Plasmodium typing results between municipal and provincial laboratories was 0.89. The Kappa values of P. falciparum, P. vivax， P. ovale and P. malaria were 0.98, 0.58, 0.87 and 0.79 respectively. Conclusion The malaria diagnosis ability of Nantong Malaria Diagnostic Reference Laboratory is generally good, and it is necessary to improve the ability of Plasmodium typing.
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Objective To re-examine the diagnosis results of reported malaria cases in Sichuan Province from 2014 to 2020, so as to assess the malaria diagnostic capability of Sichuan Provincial Malaria Diagnostic Reference Laboratory. Methods The blood and blood smear samples from reported malaria cases were collected by Sichuan Provincial Malaria Diagnostic Reference Laboratory from 2014 to 2020, and subjected to re-examinations using microscopy and nested PCR assay. The re-examination results were compared. Results A total of 1 710 samples from reported malaria cases were re-examined by Sichuan Provincial Malaria Diagnostic Reference Laboratory from 2014 to 2020, and 1 634 samples were identified positive, with a positive coincidence rate of 95.56% (1 634/1 710) and a 92.29% (1 508/1 634) total coincidence rate of the Plasmodium species. The coincidence rates with P. falciparum, P. vivax, P. malariae and P. ovale were 99.48% (961/966), 97.07% (430/443), 83.05% (98/118) and 67.86% (19/28), respectively, and the coincidence rate was 91.81% (1 513/1 648) between microscopic and nested-PCR results. Conclusions The capability of microscopists remains weak at grassroot medical institutions in Sichuan Province. Further training is required among microscopists to improve the malaria surveillance capability in Sichuan Province during the post-elimination stage.
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ObjectiveTo analyze the re-examination results of malaria cases captured from the National Notifiable Communicable Disease Reporting System in Hubei Provincial Malaria Diagnostic Reference Laboratory from 2017 to 2019, so as to pro- vide the scientific evidence for improving the malaria control capability in the province. MethodsMicroscopy and nested PCR assay were performed to re-examine the diagnosis of malaria cases registered in the National Notifiable Communicable Disease Reporting System in Hubei Provincial Malaria Diagnostic Reference Laboratory from 2017 to 2019, and the coincidences of ma- laria diagnosis and malaria parasite species were evaluated. Results A total of 410 malaria cases were reported in Hubei Province from 2017 to 2019 according to the data retrieved from the National Notifiable Communicable Disease Reporting System. Among the 407 samples re-examined by Hubei Provincial Malaria Diagnostic Reference Laboratory from 2017 to 2019, the diag- nosis 374 malaria cases were confirmed, with an overall coincidence of 91.89% (374/407) for malaria diagnosis and 89.04% (333/374) for parasite species identification. The coincidence rates of malaria diagnosis and parasite species identification were 50.00% to 100.00% and 66.67% to 100.00% in 16 cities (prefectures) of Hubei Province during the re-examinations, which both varied in regions (χ2 = 40.46 and 42.30, both P values < 0.01). The coincidence rates of Plasmodium falciparum, P. vivax, P. malariae and P. ovale identification were 95.80%, 100.00%, 58.33% and 51.92% during the re-examinations, respectively (χ2 = 76.66, P < 0.01). The consistency rate between microscopic and nested PCR results was 89.83% (362/403). Conclusions The overall diagnostic quality of malaria is high in medical institutions at all levels in Hubei Province; however, the diagnostic capability of malaria remains to be improved in some regions.
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Objective To construct the schistosomiasis diagnostic reference laboratory in Jiangsu Province, and to examine the role and diagnostic efficiency of the reference laboratory. Methods A schistosomiasis diagnostic reference laboratory was built in Jiangsu Province according to the requirements of the construction of the national schistosomiasis diagnostic reference laboratory in China. Inter-laboratory comparisons were conducted and the diagnostic capability of grassroots laboratories was evaluated in Jiangsu Province. Results The organization structure, environmental conditions, administration and quality systems of the schistosomiasis diagnostic reference laboratory in Jiangsu Province all met the requirements for construction of the national schistosomiasis diagnostic reference laboratory in China, and the schistosomiasis diagnostic reference laboratory in Jiangsu Province was issued a certificate of a province-level schistosomiasis diagnostic reference laboratory. During the 6 inter-laboratory comparisons performed by national schistosomiasis diagnostic reference centers of China, the qualitative and quantitative results of each detection item were all in agreement with the reference samples (Kappa = 1), and the diagnostic capability was identified excellent. The results of indirect hemagglutination assay of 426 serum samples from 4 grassroots laboratories were re-examined, and the mean coincidence rate was 94.13% (range, 92.08% to 96.25%) with the grassroots laboratories, with a mean Kappa value of 0.85 (range, 0.83 to 0.86) and a mean missing rate of 10.19% (range, 0 to 17.65%). Conclusions The schistosomiasis diagnostic reference laboratory has been successfully established and effectively operated in Jiangsu Province, which plays an active role in improving the capability of schistosomiasis diagnostic equality in the province.
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A three-level (provincial, municipal and county levels) schistosomiasis diagnosis network platform had been created in Yunnan Province, and assessment of laboratory quality-control samples and field evaluation of nucleic acid diagnostic techniques and immunodiagnostic reagents had been performed. This paper described the review process of the schistosomiasis diagnosis network laboratory and the operation of schistosomiasis diagnosis network laboratory and analyzed the problems of the schistosomiasis diagnosis network laboratory in Yunnan Province. The establishment of the schistosomiasis diagnosis reference (network) laboratory will provide a strong support for schistosomiasis control in Yunnan Province in the new era.
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Objective To construct the schistosomiasis diagnostic reference laboratory in Jiangsu Province, and to examine the role and diagnostic efficiency of the reference laboratory. Methods A schistosomiasis diagnostic reference laboratory was built in Jiangsu Province according to the requirements of the construction of the national schistosomiasis diagnostic reference laboratory in China. Inter-laboratory comparisons were conducted and the diagnostic capability of grassroots laboratories was evaluated in Jiangsu Province. Results The organization structure, environmental conditions, administration and quality systems of the schistosomiasis diagnostic reference laboratory in Jiangsu Province all met the requirements for construction of the national schistosomiasis diagnostic reference laboratory in China, and the schistosomiasis diagnostic reference laboratory in Jiangsu Province was issued a certificate of a province-level schistosomiasis diagnostic reference laboratory. During the 6 inter-laboratory comparisons performed by national schistosomiasis diagnostic reference centers of China, the qualitative and quantitative results of each detection item were all in agreement with the reference samples (Kappa = 1), and the diagnostic capability was identified excellent. The results of indirect hemagglutination assay of 426 serum samples from 4 grassroots laboratories were re-examined, and the mean coincidence rate was 94.13% (range, 92.08% to 96.25%) with the grassroots laboratories, with a mean Kappa value of 0.85 (range, 0.83 to 0.86) and a mean missing rate of 10.19% (range, 0 to 17.65%). Conclusions The schistosomiasis diagnostic reference laboratory has been successfully established and effectively operated in Jiangsu Province, which plays an active role in improving the capability of schistosomiasis diagnostic equality in the province.
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A three-level (provincial, municipal and county levels) schistosomiasis diagnosis network platform had been created in Yunnan Province, and assessment of laboratory quality-control samples and field evaluation of nucleic acid diagnostic techniques and immunodiagnostic reagents had been performed. This paper described the review process of the schistosomiasis diagnosis network laboratory and the operation of schistosomiasis diagnosis network laboratory and analyzed the problems of the schistosomiasis diagnosis network laboratory in Yunnan Province. The establishment of the schistosomiasis diagnosis reference (network) laboratory will provide a strong support for schistosomiasis control in Yunnan Province in the new era.
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The perpetual challenge was found for the diagnosis of newly emerging infectious diseases and reemerging infectious diseases .At present , reference laboratories for clinical microbiology were urgently demanded to deal with the challenge in China . Therefore , some suggestions for constructing reference laboratories were put forward in this paper , which may provide guidance for the future design of building comprehensive clinical microbiology laboratory .
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In this study, we report a case of anti-Gerbich (Ge) alloantibody to a high-prevalence Ge antigen in a donor with Fy(a−b−) phenotype. The alloantibody was detected in an Emirati boy who was admitted to a Korean tertiary hospital for marrow hematopoietic progenitor cell donation. He did not have a history of transfusion. His blood type was A, RhD+, and findings from the antibody screening and identification test showed 2+ reactivity in all panel cells except autologous cells. We concluded that it would be very difficult to find compatible blood components for the donor and requested further tests from external laboratories. Anti-Ge2 was identified by additional tests in a foreign reference laboratory, and the Duffy genotype of the donor was FY*02/FY*02N.01 based on the Korean Rare Blood Program. Although the donor was not a Korean, as the number of foreign patients visiting Korea increases annually, there is growing interest in patients with rare blood types in the Korean population. However, there has been very little research on rare or high prevalence blood type antigen and antibody in the Korean population. Therefore, additional research in Korea is needed on rare blood group antibodies and antigens, including Ge cases.
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Humans , Male , Antibodies , Bone Marrow , Genotype , Hematopoietic Stem Cells , Korea , Mass Screening , Phenotype , Prevalence , Tertiary Care Centers , Tissue DonorsABSTRACT
Objective To evaluate the homogeneity of the proficiency test samples to verify whether it meets the requirements of the comparison in international reference laboratory.Methods According to the Guidance on Evaluating the Homogeneity and Stability of Samples Used for Proficiency Testing (CNAS-GL03),14 biochemical indexes including ALT,AST,ALP,GGT,CK,LDH,TP,T-Bil,Urea,Cr,UA,Glu,TG and TC in the past three years (from 2014 to 2016)were tested by the Roche detection system Modular P800 Biochemical analyzer.The mean ((x)),standard deviation (s)and coefficient of variation (CV)of the samples were calculated.One-way analysis of variance (ANOVA)was performed and the guideline of Ss ≤0.3σ was used to evaluate the between-bottle differences.Results The results showed that the CVs of AST in RELA 2014A and B were higher than 2.0％.The CVs of CK were over 2％ in all tests except for RELA 2016B.The results of ANOVA for RELA samples demonstrated that the F value of CK was over the critical value 4.39,which was statistically significant (P ＜ 0.05).The F values of the ALT and T-Bil in 2015B and the Cr in 2014A were also over 4.39 (P ＜ 0.05) respectively,while the F values of other measurements were less than the critical value of F,indicating there was no statistical significance (P ＞ 0.05).The CK measurement data Ss ＞ 0.3σ in all the samples by the guideline of Ss ≤ 0.3σ,suggesting that there was a between-bottle difference in CK.The other indexes were Ss ≤ 0.3σ,showing no between-bottle difference in those items.Conclusion There were significant differences between the bottles of the CK item in the past three years,and the homogeneity of all the other items in the samples could meet the requirements of Proficiency Testing for the international reference laboratory.
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Objective To investigate the effect of measuring value transfer for human serum samples assigned by the reference laboratory network on improving the trueness of seven enzyme activities in clinical laboratories,such as ALT,AST,GGT,LDH,CK,AMY and ALP.Methods Depending on the medical imtitutions at all levels contacted by 5 reference laboratories in North China,South China,East China and Southwest China,the corresponding clinical laboratory measuring value transfer/traceability network was established.The frozen human serum samples with good interehangeability and standard material characteristics,including calibrator,sample 1 and sample 2,were provided by Beijing Aerospace General Hospital,and were assigned by 5 reference labotatories in four regiom.These samples were sent to 48 clinical laboratories.These clinical laboratories measured sample 1 and sample 2 according to their standard operating procedures,and then measured.the two samples again after adjusting their measurement system by using the supplied calibrator.The changes of trueness of detection results in these laboratories were evaluated according to the WS/T 403-2012 standard,and the changes of consistency for ALT and AST before and after measuring value tramfer were investigated.Results The results of AMY,ALP,GGT,CK and LDH calibrator,sample 1 and sample 2 assigned by the established network were 138.7 U/L,278.5 U/L and 68.3 U/L,265.3 U/L,94.5 U/L and 134.4 U/L,195.8 U/L,89.0 U/L and 158.9 U/L,393.7 U/L,260.0 U/L and 645.3 U/L,and 302.0 U/L,250.0 U/L and 452.7 U/L,respectively.The percentages of sample 1 and sample 2 which met the bias requirements of the WS/T 403-2012 standard before measuring value transfer for AMY,ALP and GGT were 65.9％ and 61.0％,76.6％ and 78.7％,and 66.7％ and 70.8％,respectively,while after measuring value transfer,they were 89.2％ and 83.8％,86.7％ and 80.0％,and 85.4％ and 91.7％,respectively.The percentages of sample 2 which met the bias requirements of the WS/T 403-2012 standard before measuring value transfer for CK and LDH were 64.6％ and 58.3％,respectively,while after measuring value trander,they were 93.5％ and 84.8％,respectively.The coefficients of variation (consistency) of sample 1 and sample 2 for ALT and AST before measuring value tramfer were 12.9％ and 11.3％,and 10.2％ and 8.9％,respectively,while after measuring value transfer,they were 9.3％ and 8.2％,and 5.6％ and 5.9％,respectively.Conclusion The calibration of routine measurement systems based on the measuring value transfer for human serum samples assigned by the reference laboratory network may improve the comparability of 7 enzyme actvities measurement results in chnical laboratories at all levels obviously,which deserves to be further spread.
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An efficient public health preparedness and response plan for infectious disease management is important in recent times when emerging and exotic diseases that hitherto were not common have surfaced in countries with potential to spread outside borders. Stewardship from a reference laboratory is important to take the lead for the laboratory network, to proactively set up disease surveillance, provide referral diagnostic services, on-going training and mentorship and to ensure coordination of an effective laboratory response. In Malaysia, the Institute for Medical Research has provided the stewardship for the Ministry of Health's laboratory network that comprises of hospital pathology, public health and university laboratories. In this paper we share our experiences in recent infectious disease outbreak investigations as a reference laboratory within the Ministry of Health infectious disease surveillance network.
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An efficient public health preparedness and response plan for infectious disease management is important in recent times when emerging and exotic diseases that hitherto were not common have surfaced in countries with potential to spread outside borders. Stewardship from a reference laboratory is important to take the lead for the laboratory network, to proactively set up disease surveillance, provide referral diagnostic services, on-going training and mentorship and to ensure coordination of an effective laboratory response. In Malaysia, the Institute for Medical Research has provided the stewardship for the Ministry of Health's laboratory network that comprises of hospital pathology, public health and university laboratories. In this paper we share our experiences in recent infectious disease outbreak investigations as a reference laboratory within the Ministry of Health infectious disease surveillance network.
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In 2010, the Japan Diabetes Society decided to introduce the National Glycohemoglobin Standardization Program (NGSP) values into clinical practice. Accordingly, NGSP Certification of Japanese manufacturers of HbA(1c)-related diagnostic reagents and instruments was initiated in February, 2012, through an NGSP network laboratory, the Asian Secondary Reference Laboratory (ASRL) #1. Traceability to the NGSP reference system can be endorsed by manufacturer certification, as well as by the College of American Pathologists (CAP) survey. Nevertheless, only a few manufacturers participate in the CAP survey in Japan. Thus, proficiency testing (PT) was proposed and executed by ASRL #1. Single-donor whole-blood samples were used for the PT. The participated measurement systems were NGSP certified. Twenty-two laboratories obtained certification through ASRL #1; 2 through the Secondary Reference Laboratory (SRL) #8; and 9 through the SRL #9. The combination plots of the bias data in this PT and in the NGSP certification performed in March and May in 2012 were consistent with each other: mean NGSP values at each level agreed well with the target value. In conclusion, PT using whole blood is useful in endorsing NGSP certification.
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Humans , Asia , Chromatography, High Pressure Liquid , Enzyme Assays , Glycated Hemoglobin/analysis , Immunoassay , Japan , Laboratory Proficiency Testing/standards , Quality Control , Reference Standards , Republic of Korea , Societies, ScientificABSTRACT
BACKGROUND: Viral screening assays performed for blood donors are required to have high sensitivity because false negative results can lead to transfusion-transmitted infections. To minimize the number of false negative cases, a systematic quality assurance program is required to verify donor screening tests. METHODS: The current status of quality assurance (QA) for blood donor screening tests in Korea and other countries was reviewed. A quality assurance program using the national standards of the Korea Food and Drug Administration (KFDA) was done as a pilot study to evaluate both the need for such a program and the feasibility of such a program. RESULTS: Singapore had a national quality assurance programs for the anti-HIVdonor screening tests. In the United Kingdom, all laboratories use the NIBSC working standards as QA materials for the donors screening. Ninety-five % (84/80) of blood centers replied that they would participate in a national quality assessment program and 92% (84/77) of the blood centers also felt that an independent organization should be designated to operate the program. Quality control materials with a weak reactivity should be included in a quality assessment program for donor screening. CONCLUSION: We propose 2 models for a National Quality Assurance Program (NQAP). In the first model, an independent national reference laboratory (NRL) needs to be established that operates the national quality assurance program. The second model involves the integration of the national quality assurance program for donor screening into the External Quality Assurance Survey run by the Korean Association of Clinical Assurance for Clinical Laboratory (KAQACL) using the national standards.
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Humans , Blood Donors , Donor Selection , United Kingdom , Korea , Mass Screening , Pilot Projects , Quality Control , Singapore , Tissue Donors , United States Food and Drug AdministrationABSTRACT
Mycobacteria that cause tuberculosis in animals and humans belong to the Mycobacterium tuberculosis complex. Techniques for conventional diagnosis are time-consuming and do not differentiate between different strains belonging to the M. tuberculosis complex. The aim of this study was to evaluate a multiplex PCR assay applicable to mycobacteria in culture with the capacity to differentiate different strains belonging to the M. tuberculosis complex in a reference laboratory. Primers based on genomics regions of difference (RD) consisting in DNA segments that are present in M. tuberculosis, but differentially deleted in several members of M. tuberculosis complex were used in a PCR assay. The test was applied to 86 clinical isolates of mycobacteria. The pattern of amplification allowed differentiating between M. tuberculosis, M. bovis and M. bovis BCG in a single PCR reaction. This PCR multiplex assay may be used in a Reference Laboratory of Tuberculosis Diagnosis as a complementary test to differentiate mycobacteria strains belonging to the M. tuberculosis complex. This test significantly reduces the time period between culture and strain identification, and thus for could favor the adoption of better strain specific antimycobacterial regimens as well as identification of zoonotic transmission of M. bovis to humans.
Las micobacterias que causan tuberculosis en animales y humanos pertenecen al complejo Mycobacterium tuberculosis. Las técnicas de diagnóstico convencional, además de ser lentas y laboriosas, no permiten diferenciar entre miembros de este complejo. El objetivo de este estudio fue evaluar ensayos de RPC múltiple para contribuir a la identificación diferencial de micobacterias del complejo M. tuberculosis a partir de cultivos, en un laboratorio de referencia. Se utilizaron oligonucleótidos partidores basados en regiones de diferencia (RD) que consisten en segmentos de ADN que están presentes en M. tuberculosis, pero que han sido eliminados diferencialmente del genoma de otros miembros del complejo M. tuberculosis. El ensayo se aplicó sobre 86 aislados clínicos de micobacterias. El patrón de amplificación permitió diferenciar entre cepas de M. tuberculosis, M. bovis y M. bovis variedad BCG en una única RPC. Este ensayo de RPC múltiple puede ser utilizado en el Laboratorio de Referencia de Diagnóstico de Tuberculosis como prueba complementaria para diferenciar micobacterias del complejo M. tuberculosis, contribuyendo a un acortamiento en el período de reporte de resultados y un tratamiento adecuado del paciente, y podría ser aplicado también en estudios epidemiológicos de transmisión zoonótica de M. bovis a humanos.