Prevalence of HIV Indeterminate Western Blot Tests and Follow-up of HIV Antibody Sero-Conversion in Southeastern China

  • Shouli WuEmail author
  • Min Gao
  • Jian Zheng
  • Pingping Yan
  • Wei Wang
  • Xiaoli Lu
  • Yuefeng Qiu
  • Yansheng YanEmail author
Research Article


HIV-indeterminate Western blotting (WB) results are typically obtained in WB confirmatory assays, and the number of indeterminate samples may increase with the detection of HIV infections, which will present considerable challenges for the management of HIV/AIDS. Nucleic acid detection has been used as a laboratory test for screening suspected or indeterminate samples. However, the effectiveness of these assays for the differential diagnosis of HIV-indeterminate WB samples remained undetermined. In this study, 210 subjects with HIV-indeterminate WB results were detected from 6360 positive HIV screening samples between 2015 and 2016 in southeastern China, in which HIV-indeterminate WB results accounted for 3.30%. The highest proportion of indeterminate results was observed in pregnant and lying-in women receiving physical examinations (16.67%), followed by that in voluntary blood donors (8.82%). The most common WB band patterns were p24, gp160 and p24, and gp160. The follow-up study revealed that the highest negative and positive conversion rates of HIV antibodies were in samples with a single p24 band (80.28%), and with gp160 and p24 bands (86.21%), respectively. Among the Env, Gag, and Pol antibodies, samples with a Gag band showed the highest negative conversion rate (81.25%), whereas the highest positive conversion rate was observed in samples with an Env band (56.76%). In addition, quantitative and qualitative HIV nucleic acid testing exhibited the highest sensitivity (96.3%) and specificity (97.85%), respectively. Our results indicate a lower proportion of HIV indeterminate WB results in southeastern China compared to previous reports, and the follow-up re-examination of patients with HIV indeterminate results should be performed. Nucleic acid testing facilitates the identification of HIV infections.


Human immunodeficiency virus (HIV) HIV indeterminate result Western blotting Nucleic acid test Differential diagnosis 



We would like to thank Drs. Hanhui Ye, Yahong Chen, and Xiaoyan Lin from the Mengchao Hepatobiliary Hospital of Fujian Medical University, and the Centers for Disease Control and Prevention in all cities of Fujian Province, for their kind assistance during the investigation. This study was supported by grants from the Cultivation of Young Talents Project Fund from the Fujian Provincial Health and Family Planning Commission (Grant No. 2015-ZQN-ZD-11), the Pilot Project of Fujian Provincial Department of Science and Technology (Grant No. 2016Y0010), and the Jiangsu Provincial Project of Invigorating Health Care through Science, Technology and Education and Jiangsu Provincial Medical Youth Talent, the Project of Invigorating Health Care through Science, Technology and Education (Grant No. QNRC2016621).

Author Contributions

SW, PY and YY designed the experiments. SW, MG and JZ carried out the experiments. SW, MG, WW, XL and YQ analyzed the data. SW, MG and WW wrote the paper. SW and YY checked and finalized the manuscript. All authors read and approved the final manuscript.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interests.

Animal and Human Rights Statement

All procedures performed in studies involving human participants were in accordance with the ethical standards of the Ethical Review Committee of Fujian Municipal Center for Disease Control and Prevention and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all participants enrolled in the study.


  1. Carneiro-Proietti AB, Cunha IW, Souza MM, Oliveira DR, Mesquita NM, Andrade CA, Catalan-Soares BC, Lima-Martins MV, Proietti FA (1999) HIV-(1/2) indeterminate western blot results: follow-up of asymptomatic blood donors in Belo Horizonte, Minas Gerais, Brazil. Rev Inst Med Trop Sao Paulo 41:155–158CrossRefGoogle Scholar
  2. CDC (2014) National HIV Testing Day and new testing recommendations. MMWR Morb Mortal Wkly Rep 63:537Google Scholar
  3. Chu CX, Zhao SP, Liu FF (2013) Analysis on follow-up outcomes of 158 samples of indeterminate HIV antibody. Lab Med Clin 10:659–660Google Scholar
  4. Coste J, Montes B, Reynes J, Peeters M, Segarra C, Vendrell JP, Delaporte E, Segondy M (1996) Comparative evaluation of three assays for the quantitation of human immunodeficiency virus type 1 RNA in plasma. J Med Virol 50:293–302CrossRefGoogle Scholar
  5. Cremonezi D, Mesquita PE, Romão MM, Prestes-Carneiro LE (2005) Prevalence of indeterminate human immunodeficiency virus western blot results in pregnant women attended at a public hospital in Presidente Prudente, Brazil. Braz J Infect Dis 9:506–509CrossRefGoogle Scholar
  6. Dodd RY, Stramer SL (2000) Indeterminate results in blood donor testing: what you don’t know can hurt you. Transfus Med Rev 14:151–160CrossRefGoogle Scholar
  7. Dou HF, Zhang HC, Chen D, Zhu CX, Wu MM, Zheng WL (2015) Investigation and analysis of the indeterminate samples of HIV antibody. Chin J Health Lab Tech 25:2973–2976Google Scholar
  8. Fauci AS (2008) 25 years of HIV. Nature 453:289–290CrossRefGoogle Scholar
  9. Fettig J, Swaminathan M, Murrill CS, Kaplan JE (2014) Global epidemiology of HIV. Infect Dis Clin North Am 28:323–337CrossRefGoogle Scholar
  10. Gao M, Wu SL, Yan YS (2016) Review of the influence factors and detection methods regarding indeterminate result of HIV antibodies by WB test. Chin J AIDS STD 22:308–311. (in Chinese)Google Scholar
  11. Granade TC, Parekh BS, Tih PM, Welty T, Welty E, Bulterys M, Ndikintum G, Nkuoh G, Tancho S (2005) Evaluation of rapid prenatal human immunodeficiency virus testing in rural cameroon. Clin Diagn Lab Immunol 12:855–860. (in Chinese)Google Scholar
  12. Guan GY, Wu ZL, Cao M, Yin T, Yang DZ, Chen H (2016) Analysis of HIV Western blot indeterminate results during 2010–2015 in Ningxia. Chin J AIDS STD 22:687–689. (in Chinese)Google Scholar
  13. Han Y, Li X, Wei ZG (2016) Analysis of indeterminate results of HIV antibodies in Xianyang City from 2010 to 2015. Chin J AIDS STD 22:831. (in Chinese)Google Scholar
  14. Jackson JB, Hanson MR, Johnson GM, Spahlinger TG, Polesky HF, Bowman RJ (1995) Long-term follow-up of blood donors with indeterminate human immunodeficiency virus type 1 results on Western blot. Transfusion 35:98–102CrossRefGoogle Scholar
  15. Jamjoom GA, Maatouk J, Gazal M, Damanhouri L, Awliaa A, Ruwaihi N, Bawazeer M, Halabi H, Adel AA, Abdulla A (1997) Follow-up of HIV western blot indeterminate results. Ann Saudi Med 17:518–521CrossRefGoogle Scholar
  16. Kakaiya R, Gordon S, Zimmerman A, Verlinsky R, Ahmed S, Phillips M (2011) False-positive nucleic acid test results for human immunodeficiency virus RNA and hepatitis C virus RNA: an underappreciated problem. Transfusion 51:225–226CrossRefGoogle Scholar
  17. Kline RL, McNairn D, Holodniy M, Mole L, Margolis D, Blattner W, Quinn TC (1996) Evaluation of Chiron HIV-1/HIV-2 recombinant immunoblot assay. J Clin Microbiol 34:2650–2653Google Scholar
  18. Liu LL, Liu BL, Xie JC, He LM (2016) Analysis of HIV-1 indeterminate Western blot results in Hechuan District, 2014 and 2015. J Dis Monit Control 10:412–413Google Scholar
  19. Maartens G, Celum C, Lewin SR (2014) HIV infection: epidemiology, pathogenesis, treatment, and prevention. Lancet 384:258–271CrossRefGoogle Scholar
  20. Mao Y, Wu Z, Poundstone K, Wang C, Qin Q, Ma Y, Ma W (2010) Development of a unified web-based national HIV/AIDS information system in China. Int J Epidemiol 39:ii79–ii89Google Scholar
  21. Miu LF, Shen YL, Wu JJ, Cheng XL, Su B (2016) Study on the serological characteristics of indeterminate HIV antibody from WB test and diagnosis supported by HIV-1 plasma viral loads. Chin J AIDS STD 22:150–154. (in Chinese)Google Scholar
  22. MOH (2016) 2015 revised National Guidelines for Detection of HIV/AIDS. Chin J Viral Dis 18:401–427. (in Chinese)Google Scholar
  23. NCAIDS, NCSTD (2017) Update on the AIDS/STD epidemic in China in February, 2017. Chin J AIDS STD 23:273Google Scholar
  24. Patel P, Mackellar D, Simmons P, Uniyal A, Gallagher K, Bennett B, Sullivan TJ, Kowalski A, Parker MM, LaLota M, Kerndt P, Sullivan PS, Centers for Disease Control and Prevention Acute HIV Infection Study Group (2010) Detecting acute human immunodeficiency virus infection using 3 different screening immunoassays and nucleic acid amplification testing for human immunodeficiency virus RNA, 2006–2008. Arch Intern Med 170:66–74CrossRefGoogle Scholar
  25. Peng JY, He JM, Jiang Y, Chen X (2014) Analysis of indeterminate results of HIV antibodies in Hunan Province from 2008 to 2012. Chin J AIDS STD 20:240–243. (in Chinese)Google Scholar
  26. SaagMS Masur H (2014) HIV/AIDS. Infect Dis Clin North Am 28:xv–xviCrossRefGoogle Scholar
  27. Schmidt M, Pichl L, Jork C, Hourfar MK, Schottstedt V, Wagner FF, Seifried E, Müller TH, Bux J, Saldanha J (2010) Blood donor screening with cobas s 201/cobasTaqScreen MPX under routine conditions at German Red Cross institutes. Vox Sang 98:37–46CrossRefGoogle Scholar
  28. Sepkowitz KA (2001) AIDS–the first 20 years. N Engl J Med 344:1764–1772CrossRefGoogle Scholar
  29. Syed IH, Balakrishnan P, Solomon SS, Murugavel KG, Kumarasamy N, Vidya S, Martin SP, Thyagarajan SP, Mayer KH, Solomon S (2005) HIV-1 western blot assay: What determines an indeterminate status? Ind J Med Sci 59:443–450CrossRefGoogle Scholar
  30. Tsoukas CM, Bernard NF (1994) Markers predicting progression of human immunodeficiency virus-related disease. Clin Microbiol Rev 7:14–28CrossRefGoogle Scholar
  31. Tuaillon E, Sanosyan A, Pisoni A, Liscouët J, Makinson A, Perre PV (2017) Staging of recent HIV-1 infection using Geenius rapid confirmatory assay compared to INNO-LIA, New Lav and Blot 2.2 assays. J Clin Virol 95:47–51CrossRefGoogle Scholar
  32. Van Gemen B, Kievits T, Nara P, Huisman HG, Jurriaans S, Goudsmit J, Lens P (1993) Qualitative and quantitative detection of HIV-1 RNA by nucleic acid sequence-based amplification. AIDS 7:S107–S110CrossRefGoogle Scholar
  33. Wang L (2007) Overview of the HIV/AIDS epidemic, scientific research and government responses in China. AIDS 21:S3–S7CrossRefGoogle Scholar
  34. Wang L, Wang Y, Jin S, Wu Z, Chin DP, Koplan JP, Wilson ME (2008) Emergence and control of infectious diseases in China. Lancet 372:1598–1605CrossRefGoogle Scholar
  35. Wei W (2015) Analysis of indeterminate results of HIV antibodies from 2008 to 2012 in Liaoyang. Chin J Health Lab Technol 25:2184–2186. (in Chinese)Google Scholar
  36. Wei M, Guan Q, Liang H, Chen JP, Chen Z, Feng Y, Hong KX, Xing H, Shao YM (2003) Development of HIV-1 laboratory diagnostic assay based on the nucleic acid detection. Chin J Lab Med 26:772–776. (in Chinese)Google Scholar
  37. Wesolowski LG, Delaney KP, Hart C, Dawson C, Owen SM, Candal D, Meyer WA 3rd, Ethridge SF, Branson BM (2011) Performance of an alternative laboratory-based algorithm for diagnosis of HIV infection utilizing a third generation immunoassay, a rapid HIV-1/HIV-2 differentiation test and a DNA or RNA-based nucleic acid amplification test in persons with established HIV-1 infection and blood donors. J ClinVirol 52:S45–S49Google Scholar
  38. World Health Organization (WHO) (2006) Patient monitoring guidelines for HIV care and antiretroviral therapy (ART). Accessed 2016-12-31
  39. Wu Z, Rou K, Cui H (2004) The HIV/AIDS epidemic in China: history, current strategies and future challenges. AIDS Educ Prev 16:7–17CrossRefGoogle Scholar
  40. Yamazaki S, Kondo M, Sudo K, Ueda T, Fujiwara H, Hasegawa N, Kato S (2016) Qualitative Real-Time PCR Assay for HIV-1 and HIV-2 RNA. Jpn J Infect Dis 69:367–372CrossRefGoogle Scholar
  41. Yan Y, Wu S, Chen L, Yan P, Qiu Y, Xie M, Wang Z, Lin X (2016) Shift in HIV/AIDS epidemic in Southeastern China: a longitudinal study from 1987 to 2015. Int J Environ Res Public Health 13:794CrossRefGoogle Scholar
  42. Yang XL, Li JY, Hao QF, Yang JM, Zhang LP, Wang L, Ma XP, Wang BH, Li J, Song X, Lu GX, Wang HY (2010) Characterization of HIV antibody indeterminate status and impact factors on HIV tests. Chin J Nosocomiol 20:1844–1846. (in Chinese)Google Scholar
  43. Zhang L, Chow EP, Jing J, Zhuang X, Li X, He M, Sun H, Li X, Gorgens M, Wilson D, Wang L, Guo W, Li D, Cui Y, Wang L, Wang N, Wu Z, Wilson DP (2013) HIV prevalence in China: integration of surveillance data and a systematic review. Lancet Infect Dis 13:955–963CrossRefGoogle Scholar
  44. Zhang XJ, Zhang L, Li X (2015) Analysis of the tracking and monitoring results in patients with uncertain HIV-1 antibody. Chin J Health Lab Tech 25:2396–2398. (in Chinese)Google Scholar
  45. Zhu M, Xiang KX, Xia XB, Hong Y, Tian F (2015) Characteristics and follow-up analysis of 27 cases with HIV antibody indeterminate result. Anhui J Prev Med 21:108–110. (in Chinese)Google Scholar

Copyright information

© Wuhan Institute of Virology, CAS 2019

Authors and Affiliations

  1. 1.Fujian Provincial Center for Disease Control and PreventionFuzhouChina
  2. 2.Fujian Provincial Key Laboratory of Zoonosis ResearchFuzhouChina
  3. 3.School of Public HealthFujian Medical UniversityFuzhouChina
  4. 4.Mengchao Hepatobiliary Hospital of Fujian Medical UniversityFuzhouChina
  5. 5.Key Laboratory of National Health Commission on Parasitic Disease Control and PreventionWuxiChina
  6. 6.Jiangsu Provincial Key Laboratory on Parasite and Vector Control TechnologyWuxiChina
  7. 7.Jiangsu Institute of Parasitic DiseasesWuxiChina

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